anti-p2x7 receptor (extracellular)-fitc antibody  (Alomone Labs)


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    Structured Review

    Alomone Labs anti-p2x7 receptor (extracellular)-fitc antibody
    Anti P2x7 Receptor (Extracellular) Fitc Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti-p2x7 receptor (extracellular)-fitc antibody/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti-p2x7 receptor (extracellular)-fitc antibody - by Bioz Stars, 2024-07
    93/100 stars

    Images

    anti p2x7 receptor extracellular fitc antibody  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
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    Structured Review

    Alomone Labs anti p2x7 receptor extracellular fitc antibody
    A) Normalized MFI of HyLite Fluor 647-labeled Aβ1-42 peptide uptake normalized to DAPI in MDMi treated with DMSO alone (Ctrl), 1 mM ATP (ATP), 1 mM ATP and 10 uM A740003 (ATP + A74), or 10 uM A740003 (A74). Batch normalization was done in GraphPad PRISM 10 with 0% defined as the smallest mean in each data set and 100% as the average of all means in the data set. Statistics were determined with a repeated measure one-way ANOVA. N=26. B) Mean Intensity of HyLite Fluor 647-labeled Aβ1-42 per cell in MDMi imaged using confocal microscopy. MDMi were treated with DMSO alone (Ctrl), 1 mM ATP (ATP), or 1 mM ATP and 10 uM A740003 (ATP + A74). Statistics were determined with a repeated measure one-way ANOVA. N= 12. C) HyLite Fluor 647-labeled Aβ1-42 peptide uptake in MDMi treated with DMSO alone (Ctrl), or 500 uM bzATP (bzATP) as measured by fluorescence on a microplate reader. MFI= Mean Fluorescence Intensity normalized to DAPI. Batch normalization was done in GraphPad PRISM 10 with 0% defined as the smallest mean in each data set and 100% as the average of all means in the data set. Statistics were determined with a paired t-test. N=14. D) Representative images of MDMi treated with DMSO alone (Ctrl) or 1 mM ATP (ATP) and stained for <t>P2RX7</t> (green) and DAPI (blue) and used to measure uptake of HyLite Fluor 647-labeled Aβ1-42 peptide (red) and were processed in CellProfiler. ns= p>0.05, *p < 0.05, ** < 0.01,*** < 0.001, **** < 0.0001
    Anti P2x7 Receptor Extracellular Fitc Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p2x7 receptor extracellular fitc antibody/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p2x7 receptor extracellular fitc antibody - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "IL-1RA Disrupts ATP Activation of P2RX7 in Human Monocyte-Derived Microglia-like Cells"

    Article Title: IL-1RA Disrupts ATP Activation of P2RX7 in Human Monocyte-Derived Microglia-like Cells

    Journal: bioRxiv

    doi: 10.1101/2024.04.08.588607

    A) Normalized MFI of HyLite Fluor 647-labeled Aβ1-42 peptide uptake normalized to DAPI in MDMi treated with DMSO alone (Ctrl), 1 mM ATP (ATP), 1 mM ATP and 10 uM A740003 (ATP + A74), or 10 uM A740003 (A74). Batch normalization was done in GraphPad PRISM 10 with 0% defined as the smallest mean in each data set and 100% as the average of all means in the data set. Statistics were determined with a repeated measure one-way ANOVA. N=26. B) Mean Intensity of HyLite Fluor 647-labeled Aβ1-42 per cell in MDMi imaged using confocal microscopy. MDMi were treated with DMSO alone (Ctrl), 1 mM ATP (ATP), or 1 mM ATP and 10 uM A740003 (ATP + A74). Statistics were determined with a repeated measure one-way ANOVA. N= 12. C) HyLite Fluor 647-labeled Aβ1-42 peptide uptake in MDMi treated with DMSO alone (Ctrl), or 500 uM bzATP (bzATP) as measured by fluorescence on a microplate reader. MFI= Mean Fluorescence Intensity normalized to DAPI. Batch normalization was done in GraphPad PRISM 10 with 0% defined as the smallest mean in each data set and 100% as the average of all means in the data set. Statistics were determined with a paired t-test. N=14. D) Representative images of MDMi treated with DMSO alone (Ctrl) or 1 mM ATP (ATP) and stained for P2RX7 (green) and DAPI (blue) and used to measure uptake of HyLite Fluor 647-labeled Aβ1-42 peptide (red) and were processed in CellProfiler. ns= p>0.05, *p < 0.05, ** < 0.01,*** < 0.001, **** < 0.0001
    Figure Legend Snippet: A) Normalized MFI of HyLite Fluor 647-labeled Aβ1-42 peptide uptake normalized to DAPI in MDMi treated with DMSO alone (Ctrl), 1 mM ATP (ATP), 1 mM ATP and 10 uM A740003 (ATP + A74), or 10 uM A740003 (A74). Batch normalization was done in GraphPad PRISM 10 with 0% defined as the smallest mean in each data set and 100% as the average of all means in the data set. Statistics were determined with a repeated measure one-way ANOVA. N=26. B) Mean Intensity of HyLite Fluor 647-labeled Aβ1-42 per cell in MDMi imaged using confocal microscopy. MDMi were treated with DMSO alone (Ctrl), 1 mM ATP (ATP), or 1 mM ATP and 10 uM A740003 (ATP + A74). Statistics were determined with a repeated measure one-way ANOVA. N= 12. C) HyLite Fluor 647-labeled Aβ1-42 peptide uptake in MDMi treated with DMSO alone (Ctrl), or 500 uM bzATP (bzATP) as measured by fluorescence on a microplate reader. MFI= Mean Fluorescence Intensity normalized to DAPI. Batch normalization was done in GraphPad PRISM 10 with 0% defined as the smallest mean in each data set and 100% as the average of all means in the data set. Statistics were determined with a paired t-test. N=14. D) Representative images of MDMi treated with DMSO alone (Ctrl) or 1 mM ATP (ATP) and stained for P2RX7 (green) and DAPI (blue) and used to measure uptake of HyLite Fluor 647-labeled Aβ1-42 peptide (red) and were processed in CellProfiler. ns= p>0.05, *p < 0.05, ** < 0.01,*** < 0.001, **** < 0.0001

    Techniques Used: Labeling, Confocal Microscopy, Fluorescence, Staining

    rabbit anti p2x7r monoclonal igg  (Alomone Labs)


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  • 93

    Structured Review

    Alomone Labs rabbit anti p2x7r monoclonal igg
    The sequences of oligonucleotide primers.
    Rabbit Anti P2x7r Monoclonal Igg, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti p2x7r monoclonal igg/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti p2x7r monoclonal igg - by Bioz Stars, 2024-07
    93/100 stars

    Images

    1) Product Images from "The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain"

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    Journal: BioMed Research International

    doi: 10.1155/2020/8143754

    The sequences of oligonucleotide primers.
    Figure Legend Snippet: The sequences of oligonucleotide primers.

    Techniques Used:

    The antibodies for immunoblotting.
    Figure Legend Snippet: The antibodies for immunoblotting.

    Techniques Used: Western Blot, Concentration Assay

    The antibodies for fluorescence labeling.
    Figure Legend Snippet: The antibodies for fluorescence labeling.

    Techniques Used: Fluorescence, Labeling, Concentration Assay

    Mechanical threshold and P2X7R and p-p38 expression. (a) Mechanical threshold after BTZ injection. (b, c) Western blot for P2X7R expression after BTZ treatment. (d) Immunofluorescence location of P2X7R in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. P2X7R is not expressed in NF-200-positive neurons. P2X7R is expressed in GFAP-labeled satellite glial cells (SGCs). (e) Immunofluorescence location of p-p38 in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. p-p38 is expressed in both MAP2-labeled neurons and GFAP-labeled SGCs. (f) Immunofluorescence location of P2X7R in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. P2X7R is expressed mainly in Iba-1-labeled microglial cells rather than in GFAP-labeled astrocytes and MAP2-labeled neurons. (g) Immunofluorescence location of p-p38 in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. p-p38 is expressed mainly in Iba-1-labeled microglial cells. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: Mechanical threshold and P2X7R and p-p38 expression. (a) Mechanical threshold after BTZ injection. (b, c) Western blot for P2X7R expression after BTZ treatment. (d) Immunofluorescence location of P2X7R in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. P2X7R is not expressed in NF-200-positive neurons. P2X7R is expressed in GFAP-labeled satellite glial cells (SGCs). (e) Immunofluorescence location of p-p38 in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. p-p38 is expressed in both MAP2-labeled neurons and GFAP-labeled SGCs. (f) Immunofluorescence location of P2X7R in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. P2X7R is expressed mainly in Iba-1-labeled microglial cells rather than in GFAP-labeled astrocytes and MAP2-labeled neurons. (g) Immunofluorescence location of p-p38 in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. p-p38 is expressed mainly in Iba-1-labeled microglial cells. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Injection, Western Blot, Immunofluorescence, Labeling

    p38 mRNA expression and p38 phosphorylation in DRG after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) p-p38 immunofluorescence labeling. The arrows show the typical p-p38 single-labeled DRG cells. (e) p-p38 fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗∗ P < 0.001.
    Figure Legend Snippet: p38 mRNA expression and p38 phosphorylation in DRG after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) p-p38 immunofluorescence labeling. The arrows show the typical p-p38 single-labeled DRG cells. (e) p-p38 fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Immunofluorescence, Labeling, Fluorescence

    p38 mRNA expression and p38 phosphorylation in SDH after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: p38 mRNA expression and p38 phosphorylation in SDH after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    IL-1 β , IL-6, and TNF- α mRNA expression in DRG and SDH after inhibition of P2X7R. (a) DRG IL-1 β mRNA. (b) DRG IL-6 mRNA. (c) DRG TNF- α mRNA. (d) SDH IL-1 β mRNA. (e) SDH IL-6 mRNA. (f) SDH TNF- α mRNA. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: IL-1 β , IL-6, and TNF- α mRNA expression in DRG and SDH after inhibition of P2X7R. (a) DRG IL-1 β mRNA. (b) DRG IL-6 mRNA. (c) DRG TNF- α mRNA. (d) SDH IL-1 β mRNA. (e) SDH IL-6 mRNA. (f) SDH TNF- α mRNA. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition

    P2X7R mRNA and protein expression in DRG after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and GFAP coexpression fluorescence labeling for SGCs. The arrows indicate the typical single-labeled and double-labeled DRG satellite cells. (e) P2X7R and GFAP coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: P2X7R mRNA and protein expression in DRG after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and GFAP coexpression fluorescence labeling for SGCs. The arrows indicate the typical single-labeled and double-labeled DRG satellite cells. (e) P2X7R and GFAP coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    P2X7R mRNA and protein expression in SDH after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 colocalization fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: P2X7R mRNA and protein expression in SDH after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 colocalization fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    Mechanical threshold alterations after inhibition of P2X7R or p38. (a) Mechanical threshold after inhibition of P2X7R. (b) Mechanical threshold after inhibition of p38. Mean ± SEM ( n = 5). ∗∗∗ P < 0.001 (vs. control); # P < 0.05; ## P < 0.01 (vs. BTZ group).
    Figure Legend Snippet: Mechanical threshold alterations after inhibition of P2X7R or p38. (a) Mechanical threshold after inhibition of P2X7R. (b) Mechanical threshold after inhibition of p38. Mean ± SEM ( n = 5). ∗∗∗ P < 0.001 (vs. control); # P < 0.05; ## P < 0.01 (vs. BTZ group).

    Techniques Used: Inhibition

    rabbit anti p2x7r monoclonal igg  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
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    Structured Review

    Alomone Labs rabbit anti p2x7r monoclonal igg
    The sequences of oligonucleotide primers.
    Rabbit Anti P2x7r Monoclonal Igg, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti p2x7r monoclonal igg/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti p2x7r monoclonal igg - by Bioz Stars, 2024-07
    93/100 stars

    Images

    1) Product Images from "The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain"

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    Journal: BioMed Research International

    doi: 10.1155/2020/8143754

    The sequences of oligonucleotide primers.
    Figure Legend Snippet: The sequences of oligonucleotide primers.

    Techniques Used:

    The antibodies for immunoblotting.
    Figure Legend Snippet: The antibodies for immunoblotting.

    Techniques Used: Western Blot, Concentration Assay

    The antibodies for fluorescence labeling.
    Figure Legend Snippet: The antibodies for fluorescence labeling.

    Techniques Used: Fluorescence, Labeling, Concentration Assay

    Mechanical threshold and P2X7R and p-p38 expression. (a) Mechanical threshold after BTZ injection. (b, c) Western blot for P2X7R expression after BTZ treatment. (d) Immunofluorescence location of P2X7R in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. P2X7R is not expressed in NF-200-positive neurons. P2X7R is expressed in GFAP-labeled satellite glial cells (SGCs). (e) Immunofluorescence location of p-p38 in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. p-p38 is expressed in both MAP2-labeled neurons and GFAP-labeled SGCs. (f) Immunofluorescence location of P2X7R in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. P2X7R is expressed mainly in Iba-1-labeled microglial cells rather than in GFAP-labeled astrocytes and MAP2-labeled neurons. (g) Immunofluorescence location of p-p38 in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. p-p38 is expressed mainly in Iba-1-labeled microglial cells. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: Mechanical threshold and P2X7R and p-p38 expression. (a) Mechanical threshold after BTZ injection. (b, c) Western blot for P2X7R expression after BTZ treatment. (d) Immunofluorescence location of P2X7R in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. P2X7R is not expressed in NF-200-positive neurons. P2X7R is expressed in GFAP-labeled satellite glial cells (SGCs). (e) Immunofluorescence location of p-p38 in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. p-p38 is expressed in both MAP2-labeled neurons and GFAP-labeled SGCs. (f) Immunofluorescence location of P2X7R in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. P2X7R is expressed mainly in Iba-1-labeled microglial cells rather than in GFAP-labeled astrocytes and MAP2-labeled neurons. (g) Immunofluorescence location of p-p38 in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. p-p38 is expressed mainly in Iba-1-labeled microglial cells. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Injection, Western Blot, Immunofluorescence, Labeling

    p38 mRNA expression and p38 phosphorylation in DRG after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) p-p38 immunofluorescence labeling. The arrows show the typical p-p38 single-labeled DRG cells. (e) p-p38 fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗∗ P < 0.001.
    Figure Legend Snippet: p38 mRNA expression and p38 phosphorylation in DRG after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) p-p38 immunofluorescence labeling. The arrows show the typical p-p38 single-labeled DRG cells. (e) p-p38 fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Immunofluorescence, Labeling, Fluorescence

    p38 mRNA expression and p38 phosphorylation in SDH after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: p38 mRNA expression and p38 phosphorylation in SDH after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    IL-1 β , IL-6, and TNF- α mRNA expression in DRG and SDH after inhibition of P2X7R. (a) DRG IL-1 β mRNA. (b) DRG IL-6 mRNA. (c) DRG TNF- α mRNA. (d) SDH IL-1 β mRNA. (e) SDH IL-6 mRNA. (f) SDH TNF- α mRNA. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: IL-1 β , IL-6, and TNF- α mRNA expression in DRG and SDH after inhibition of P2X7R. (a) DRG IL-1 β mRNA. (b) DRG IL-6 mRNA. (c) DRG TNF- α mRNA. (d) SDH IL-1 β mRNA. (e) SDH IL-6 mRNA. (f) SDH TNF- α mRNA. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition

    P2X7R mRNA and protein expression in DRG after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and GFAP coexpression fluorescence labeling for SGCs. The arrows indicate the typical single-labeled and double-labeled DRG satellite cells. (e) P2X7R and GFAP coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: P2X7R mRNA and protein expression in DRG after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and GFAP coexpression fluorescence labeling for SGCs. The arrows indicate the typical single-labeled and double-labeled DRG satellite cells. (e) P2X7R and GFAP coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    P2X7R mRNA and protein expression in SDH after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 colocalization fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.
    Figure Legend Snippet: P2X7R mRNA and protein expression in SDH after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 colocalization fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Techniques Used: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    Mechanical threshold alterations after inhibition of P2X7R or p38. (a) Mechanical threshold after inhibition of P2X7R. (b) Mechanical threshold after inhibition of p38. Mean ± SEM ( n = 5). ∗∗∗ P < 0.001 (vs. control); # P < 0.05; ## P < 0.01 (vs. BTZ group).
    Figure Legend Snippet: Mechanical threshold alterations after inhibition of P2X7R or p38. (a) Mechanical threshold after inhibition of P2X7R. (b) Mechanical threshold after inhibition of p38. Mean ± SEM ( n = 5). ∗∗∗ P < 0.001 (vs. control); # P < 0.05; ## P < 0.01 (vs. BTZ group).

    Techniques Used: Inhibition

    anti p2x7 receptor extracellular fitc antibody  (Alomone Labs)


    Bioz Verified Symbol Alomone Labs is a verified supplier
    Bioz Manufacturer Symbol Alomone Labs manufactures this product  
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  • 93

    Structured Review

    Alomone Labs anti p2x7 receptor extracellular fitc antibody
    <t>P2X7R</t> regulates the development of acute gouty arthritis in rats. ( A ) At 12h, clinical manifestations of right ankle joint in rats among ATP group, BBG group and control group. ( B and C ) Clinical score and swelling index of three groups of rats at each time point. ( D ) Inflammatory cells infiltrated the synovial tissue of the right ankle joint in three groups of rats, at 24h. ( E and F ) Infiltration of mononuclear cells and neutrophils in the synovial tissue of the ankle joint among the three groups. * P<0.05, **P< 0.01, ***P< 0.001.
    Anti P2x7 Receptor Extracellular Fitc Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti p2x7 receptor extracellular fitc antibody/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti p2x7 receptor extracellular fitc antibody - by Bioz Stars, 2024-07
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    1) Product Images from "ATP-Activated P2X7R Promote the Attack of Acute Gouty Arthritis in Rats Through Activating NLRP3 Inflammasome and Inflammatory Cytokine Production"

    Article Title: ATP-Activated P2X7R Promote the Attack of Acute Gouty Arthritis in Rats Through Activating NLRP3 Inflammasome and Inflammatory Cytokine Production

    Journal: Journal of Inflammation Research

    doi: 10.2147/JIR.S351660

    P2X7R regulates the development of acute gouty arthritis in rats. ( A ) At 12h, clinical manifestations of right ankle joint in rats among ATP group, BBG group and control group. ( B and C ) Clinical score and swelling index of three groups of rats at each time point. ( D ) Inflammatory cells infiltrated the synovial tissue of the right ankle joint in three groups of rats, at 24h. ( E and F ) Infiltration of mononuclear cells and neutrophils in the synovial tissue of the ankle joint among the three groups. * P<0.05, **P< 0.01, ***P< 0.001.
    Figure Legend Snippet: P2X7R regulates the development of acute gouty arthritis in rats. ( A ) At 12h, clinical manifestations of right ankle joint in rats among ATP group, BBG group and control group. ( B and C ) Clinical score and swelling index of three groups of rats at each time point. ( D ) Inflammatory cells infiltrated the synovial tissue of the right ankle joint in three groups of rats, at 24h. ( E and F ) Infiltration of mononuclear cells and neutrophils in the synovial tissue of the ankle joint among the three groups. * P<0.05, **P< 0.01, ***P< 0.001.

    Techniques Used:

    The expression changes of P2X7R on macrophages affect their ability to take up YO-PRO-1. ( A ) At 12h, the expression level of P2X7R in each group of macrophages was detected by flow cytometry. ( B ) Expression of P2X7R among the three groups at different time points. ( C ) At 12h, the percentage of YO-PRO-1 uptake by macrophages in each group was detected by flow cytometry. ( D ) The ability of macrophages to uptake YO-PRO-1 among the three groups at each time point. ( E ) The expression level of P2X7R was positively correlated with the ability of macrophages to uptake YO-PRO-1. * P<0.05, ** P< 0.01, *** P< 0.001.
    Figure Legend Snippet: The expression changes of P2X7R on macrophages affect their ability to take up YO-PRO-1. ( A ) At 12h, the expression level of P2X7R in each group of macrophages was detected by flow cytometry. ( B ) Expression of P2X7R among the three groups at different time points. ( C ) At 12h, the percentage of YO-PRO-1 uptake by macrophages in each group was detected by flow cytometry. ( D ) The ability of macrophages to uptake YO-PRO-1 among the three groups at each time point. ( E ) The expression level of P2X7R was positively correlated with the ability of macrophages to uptake YO-PRO-1. * P<0.05, ** P< 0.01, *** P< 0.001.

    Techniques Used: Expressing, Flow Cytometry

    P2X7R mediated NLRP3 inflammatory-dependent IL-1β secretion on macrophages. The expressions of P2X7R ( A ), NLRP3 ( B ) and IL-1β ( C ) in rat spleen macrophages were analyzed by qRT-PCR. *P<0.05, **P< 0.01, ***P< 0.001.
    Figure Legend Snippet: P2X7R mediated NLRP3 inflammatory-dependent IL-1β secretion on macrophages. The expressions of P2X7R ( A ), NLRP3 ( B ) and IL-1β ( C ) in rat spleen macrophages were analyzed by qRT-PCR. *P<0.05, **P< 0.01, ***P< 0.001.

    Techniques Used: Quantitative RT-PCR

    P2X7R promote Th17 cells and pro-inflammatory cytokines production. ( A ) The expressions of CD4 + IL-17 + Th17 cells among the three groups at 12h. ( B ) The levels of CD4 + IL-17 + Th17 cells in ATP group, BBG group and Control group at each time point. ( C-E ) The serum levels of IL-1β, IL-6 and IL-17 of the three groups at each time point. (F) The expression of IL-17 mRNA among three groups at each time point. * P<0.05, ** P< 0.01, *** P< 0.001.
    Figure Legend Snippet: P2X7R promote Th17 cells and pro-inflammatory cytokines production. ( A ) The expressions of CD4 + IL-17 + Th17 cells among the three groups at 12h. ( B ) The levels of CD4 + IL-17 + Th17 cells in ATP group, BBG group and Control group at each time point. ( C-E ) The serum levels of IL-1β, IL-6 and IL-17 of the three groups at each time point. (F) The expression of IL-17 mRNA among three groups at each time point. * P<0.05, ** P< 0.01, *** P< 0.001.

    Techniques Used: Expressing

    Tregs and anti-inflammatory cytokines in P2X7R-regulated acute gouty arthritis. ( A ) The expressions of Tregs among the three groups at 12h. ( B ) The levels of CD4 + CD25 + FOXP3 + Tregs in three groups at each time point. ( C) The expression of FOXP3 mRNA among three groups at each time point. ( D -E) The expressions of TGF-β 1 and IL-10 in serum among the three groups. *P<0.05, **P< 0.01, ***P< 0.001.
    Figure Legend Snippet: Tregs and anti-inflammatory cytokines in P2X7R-regulated acute gouty arthritis. ( A ) The expressions of Tregs among the three groups at 12h. ( B ) The levels of CD4 + CD25 + FOXP3 + Tregs in three groups at each time point. ( C) The expression of FOXP3 mRNA among three groups at each time point. ( D -E) The expressions of TGF-β 1 and IL-10 in serum among the three groups. *P<0.05, **P< 0.01, ***P< 0.001.

    Techniques Used: Expressing

    The changing trend of cytokines and Treg/Th17 cells matched with the pathogenesis process of ATP-activated P2X7R-regulated acute gouty arthritis. ( A ) The trends of P2X7R and IL-1β in ATP group. ( B ) The trends of CD4 + IL-17 + Th17 cells and IL-17 in ATP group. ( C ) The trends of CD4 + CD25 + Foxp3 + Treg cells and TGF-β 1 in ATP group. ( D ) The trends of CD4 + CD25 + Foxp3 + Treg cells and CD4 + IL-17 + Th17 cells in ATP group. ( E ) The ratios of Treg/Th17 among ATP group, BBG group and Control group. * P<0.05, ATP group vs BBG group, # P<0.05, ATP group vs Control group.
    Figure Legend Snippet: The changing trend of cytokines and Treg/Th17 cells matched with the pathogenesis process of ATP-activated P2X7R-regulated acute gouty arthritis. ( A ) The trends of P2X7R and IL-1β in ATP group. ( B ) The trends of CD4 + IL-17 + Th17 cells and IL-17 in ATP group. ( C ) The trends of CD4 + CD25 + Foxp3 + Treg cells and TGF-β 1 in ATP group. ( D ) The trends of CD4 + CD25 + Foxp3 + Treg cells and CD4 + IL-17 + Th17 cells in ATP group. ( E ) The ratios of Treg/Th17 among ATP group, BBG group and Control group. * P<0.05, ATP group vs BBG group, # P<0.05, ATP group vs Control group.

    Techniques Used:

    fluorescein isothiocyanate fitc  (Alomone Labs)


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    Alomone Labs fluorescein isothiocyanate fitc
    Fluorescein Isothiocyanate Fitc, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    antibodies anti p2x7 fitc  (Alomone Labs)


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    Alomone Labs antibodies anti p2x7 fitc
    <t>P2X7</t> receptors identified by immunofluorescence, in renal cortex and medulla in Sham and Ang II-infused rats. The expression of P2X7 receptors in the Ang II group is localized in tubular membranes ( A ). Western blot ( B ) showed abundance of the P2X7 receptor protein in the Ang II group ( n = 7 per group). * p < 0.01 vs. Sham. Ang II = angiotensin II, BBG = Brilliant blue G.
    Antibodies Anti P2x7 Fitc, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "The Role of P2X7 Purinergic Receptors in the Renal Inflammation Associated with Angiotensin II-Induced Hypertension"

    Article Title: The Role of P2X7 Purinergic Receptors in the Renal Inflammation Associated with Angiotensin II-Induced Hypertension

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms21114041

    P2X7 receptors identified by immunofluorescence, in renal cortex and medulla in Sham and Ang II-infused rats. The expression of P2X7 receptors in the Ang II group is localized in tubular membranes ( A ). Western blot ( B ) showed abundance of the P2X7 receptor protein in the Ang II group ( n = 7 per group). * p < 0.01 vs. Sham. Ang II = angiotensin II, BBG = Brilliant blue G.
    Figure Legend Snippet: P2X7 receptors identified by immunofluorescence, in renal cortex and medulla in Sham and Ang II-infused rats. The expression of P2X7 receptors in the Ang II group is localized in tubular membranes ( A ). Western blot ( B ) showed abundance of the P2X7 receptor protein in the Ang II group ( n = 7 per group). * p < 0.01 vs. Sham. Ang II = angiotensin II, BBG = Brilliant blue G.

    Techniques Used: Immunofluorescence, Expressing, Western Blot

    P2X7 receptors (second column) in the infiltrating T cells of Ang II group co-localize with cluster differentiation (CD, first column) antigens in T cells (CD 3 positive cells), B cells (CD5 and CD20 positive cells), macrophages (CD68 positive cells) and leukocyte common antigen CD45, predominantly expressed in T lymphocytes.
    Figure Legend Snippet: P2X7 receptors (second column) in the infiltrating T cells of Ang II group co-localize with cluster differentiation (CD, first column) antigens in T cells (CD 3 positive cells), B cells (CD5 and CD20 positive cells), macrophages (CD68 positive cells) and leukocyte common antigen CD45, predominantly expressed in T lymphocytes.

    Techniques Used:

    anti-p2x7 receptor (extracellular)-fitc antibody  (Alomone Labs)


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    Alomone Labs anti-p2x7 receptor (extracellular)-fitc antibody
    Anti P2x7 Receptor (Extracellular) Fitc Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    p2x7r antibody incubation  (Alomone Labs)


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    Alomone Labs p2x7r antibody incubation
    P2x7r Antibody Incubation, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    p2x7r antibody incubation  (Alomone Labs)


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    Alomone Labs p2x7r antibody incubation
    P2x7r Antibody Incubation, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti p2x7r extracellular fitc  (Alomone Labs)


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    Alomone Labs anti p2x7r extracellular fitc
    P24XR and <t>P2X7R</t> Mediate ATP-Dependent Calcium Signal Propagation (A and B) Murine RAW 264.7 macrophages were loaded with photoactivable caged-IP 3 and the fluorescent calcium indicator Fluo-4 (green), and calcium signal propagation after IP 3 uncaging in the origin cell (white box) was monitored in live imaging. Experiments were performed in HBSS with 2 mM Ca 2+ (A) or in calcium-free HBSS supplemented with 2 mM EGTA (B). Scale bars, 50 μm. See also . (C and D) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 (C) or primary BMDMs (D). Error bars represent SEM. For statistical data analysis, Student’s t test was used ( ∗ p < 0.05). (E and F) Relative mRNA expression of different members of the P2X family of receptors, measured by real-time PCR in the RAW 264.7 cell line (E) and in BMDM primary macrophages (F). Error bars represent SEM. (G) Quantification of 5 independent live calcium imaging experiments with RAW 264.7 cells pre-treated with the P2X4R inhibitor 5BDBD (100 μM, 30 min at 37°C), the P2X7R inhibitor A740003 (100 μM, 30 min at 37°C), or their vehicle (DMSO) or left untreated. Error bars represent SEM. For statistical data analysis, One-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant). (H) Representative western blot (left) and quantification of repeated experiments (right) of P2X4R and P2X7R expression in RAW 264.7 cells transfected with siRNA specific for P2X4R and P2X7R or with scramble siRNA. Control cells were electroporated in the absence of oligonucleotides. (I) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 cells silenced for P2X4R and P2X7R. Error bars represent SEM. For statistical data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant).
    Anti P2x7r Extracellular Fitc, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Intercellular Calcium Signaling Induced by ATP Potentiates Macrophage Phagocytosis"

    Article Title: Intercellular Calcium Signaling Induced by ATP Potentiates Macrophage Phagocytosis

    Journal: Cell Reports

    doi: 10.1016/j.celrep.2019.03.011

    P24XR and P2X7R Mediate ATP-Dependent Calcium Signal Propagation (A and B) Murine RAW 264.7 macrophages were loaded with photoactivable caged-IP 3 and the fluorescent calcium indicator Fluo-4 (green), and calcium signal propagation after IP 3 uncaging in the origin cell (white box) was monitored in live imaging. Experiments were performed in HBSS with 2 mM Ca 2+ (A) or in calcium-free HBSS supplemented with 2 mM EGTA (B). Scale bars, 50 μm. See also . (C and D) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 (C) or primary BMDMs (D). Error bars represent SEM. For statistical data analysis, Student’s t test was used ( ∗ p < 0.05). (E and F) Relative mRNA expression of different members of the P2X family of receptors, measured by real-time PCR in the RAW 264.7 cell line (E) and in BMDM primary macrophages (F). Error bars represent SEM. (G) Quantification of 5 independent live calcium imaging experiments with RAW 264.7 cells pre-treated with the P2X4R inhibitor 5BDBD (100 μM, 30 min at 37°C), the P2X7R inhibitor A740003 (100 μM, 30 min at 37°C), or their vehicle (DMSO) or left untreated. Error bars represent SEM. For statistical data analysis, One-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant). (H) Representative western blot (left) and quantification of repeated experiments (right) of P2X4R and P2X7R expression in RAW 264.7 cells transfected with siRNA specific for P2X4R and P2X7R or with scramble siRNA. Control cells were electroporated in the absence of oligonucleotides. (I) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 cells silenced for P2X4R and P2X7R. Error bars represent SEM. For statistical data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant).
    Figure Legend Snippet: P24XR and P2X7R Mediate ATP-Dependent Calcium Signal Propagation (A and B) Murine RAW 264.7 macrophages were loaded with photoactivable caged-IP 3 and the fluorescent calcium indicator Fluo-4 (green), and calcium signal propagation after IP 3 uncaging in the origin cell (white box) was monitored in live imaging. Experiments were performed in HBSS with 2 mM Ca 2+ (A) or in calcium-free HBSS supplemented with 2 mM EGTA (B). Scale bars, 50 μm. See also . (C and D) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 (C) or primary BMDMs (D). Error bars represent SEM. For statistical data analysis, Student’s t test was used ( ∗ p < 0.05). (E and F) Relative mRNA expression of different members of the P2X family of receptors, measured by real-time PCR in the RAW 264.7 cell line (E) and in BMDM primary macrophages (F). Error bars represent SEM. (G) Quantification of 5 independent live calcium imaging experiments with RAW 264.7 cells pre-treated with the P2X4R inhibitor 5BDBD (100 μM, 30 min at 37°C), the P2X7R inhibitor A740003 (100 μM, 30 min at 37°C), or their vehicle (DMSO) or left untreated. Error bars represent SEM. For statistical data analysis, One-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant). (H) Representative western blot (left) and quantification of repeated experiments (right) of P2X4R and P2X7R expression in RAW 264.7 cells transfected with siRNA specific for P2X4R and P2X7R or with scramble siRNA. Control cells were electroporated in the absence of oligonucleotides. (I) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 cells silenced for P2X4R and P2X7R. Error bars represent SEM. For statistical data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant).

    Techniques Used: Imaging, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Transfection

    Macrophage Polarization Status Affects Calcium Signal Propagation (A) The surface expression of P2X4R (top) and P2X7R (bottom) was analyzed by flow cytometry in resting, IFNγ-treated (10 ng/mL, 24 h), or IL4-treated (20 ng/mL, 24 h) macrophages. Histograms show the quantification 3 independent biological replicates. Error bars represent SEM. For data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used (ns, non-significant; ∗∗ p < 0.01; ∗∗∗ p < 0.001). (B) Maximal back-projection of two representative live calcium imaging experiments performed with IFNγ- or IL4-treated RAW 264.7 cells loaded with caged-IP 3 and Fluo-4. The fluorescence variation 60 s after the irradiation of the origin cell (white box) is represented in false colors. (C) Representative traces of live calcium imaging experiments, showing the fluorescence variation after the uncaging of the origin cell (red) and the bystander macrophages (black). (D) Quantification of 3 independent live calcium imaging experiments. Error bars represent SEM. For data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant).
    Figure Legend Snippet: Macrophage Polarization Status Affects Calcium Signal Propagation (A) The surface expression of P2X4R (top) and P2X7R (bottom) was analyzed by flow cytometry in resting, IFNγ-treated (10 ng/mL, 24 h), or IL4-treated (20 ng/mL, 24 h) macrophages. Histograms show the quantification 3 independent biological replicates. Error bars represent SEM. For data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used (ns, non-significant; ∗∗ p < 0.01; ∗∗∗ p < 0.001). (B) Maximal back-projection of two representative live calcium imaging experiments performed with IFNγ- or IL4-treated RAW 264.7 cells loaded with caged-IP 3 and Fluo-4. The fluorescence variation 60 s after the irradiation of the origin cell (white box) is represented in false colors. (C) Representative traces of live calcium imaging experiments, showing the fluorescence variation after the uncaging of the origin cell (red) and the bystander macrophages (black). (D) Quantification of 3 independent live calcium imaging experiments. Error bars represent SEM. For data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant).

    Techniques Used: Expressing, Flow Cytometry, Imaging, Fluorescence, Irradiation

    Extracellular ATP Is Required for Efficient Phagocytosis (A) Primary BMDMs were incubated with PhRodo E. coli fluorescent bioparticles in the presence or absence of 5 mM EGTA to chelate extracellular calcium. Phagocytosis was monitored at 15 or 30 min by flow cytometry (see <xref ref-type=Figure S4 ). Macrophages incubated with 20 μM cytochalasin D were used as negative reference. The phagocytic index was calculated as the percentage of fluorescent macrophages multiplied by their mean of fluorescence (MFI) and normalized on the cytochalasin-treated samples. (B) Primary BMDMs were loaded with the intracellular calcium chelator BAPTA-AM or its vehicle (loading solution) before performing the phagocytosis assay. (C) Primary BMDMs were incubated with PhRodo E. coli , PhRodo Zymosan, or PhRodo S. aureus fluorescent bioparticles, in the presence or absence of apyrase (5 U/mL). (D) Primary BMDMs were pretreated with the P2X4R inhibitor 5BDBD (100 μM), the P2X7R inhibitor A740003 (100 μM), or their vehicle (DMSO), or were left untreated, before performing the phagocytosis assay. (E) Phagocytosis was performed for 30 min in the presence or absence of MSC-derived EVs, pre-incubated or not with ARL-67516 (30 min, 200 μM). The graphs are representative of at least 3 independent biological replicates, each performed in technical triplicate. Error bars represent SEM. For data analysis, a two-way ANOVA followed by Tukey’s multiple comparisons test was used ( ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, non-significant). " title="... with the P2X4R inhibitor 5BDBD (100 μM), the P2X7R inhibitor A740003 (100 μM), or their vehicle (DMSO), ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Extracellular ATP Is Required for Efficient Phagocytosis (A) Primary BMDMs were incubated with PhRodo E. coli fluorescent bioparticles in the presence or absence of 5 mM EGTA to chelate extracellular calcium. Phagocytosis was monitored at 15 or 30 min by flow cytometry (see Figure S4 ). Macrophages incubated with 20 μM cytochalasin D were used as negative reference. The phagocytic index was calculated as the percentage of fluorescent macrophages multiplied by their mean of fluorescence (MFI) and normalized on the cytochalasin-treated samples. (B) Primary BMDMs were loaded with the intracellular calcium chelator BAPTA-AM or its vehicle (loading solution) before performing the phagocytosis assay. (C) Primary BMDMs were incubated with PhRodo E. coli , PhRodo Zymosan, or PhRodo S. aureus fluorescent bioparticles, in the presence or absence of apyrase (5 U/mL). (D) Primary BMDMs were pretreated with the P2X4R inhibitor 5BDBD (100 μM), the P2X7R inhibitor A740003 (100 μM), or their vehicle (DMSO), or were left untreated, before performing the phagocytosis assay. (E) Phagocytosis was performed for 30 min in the presence or absence of MSC-derived EVs, pre-incubated or not with ARL-67516 (30 min, 200 μM). The graphs are representative of at least 3 independent biological replicates, each performed in technical triplicate. Error bars represent SEM. For data analysis, a two-way ANOVA followed by Tukey’s multiple comparisons test was used ( ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, non-significant).

    Techniques Used: Incubation, Flow Cytometry, Fluorescence, Phagocytosis Assay, Derivative Assay


    Figure Legend Snippet:

    Techniques Used: Recombinant, Negative Control, Real-time Polymerase Chain Reaction, Software


    Figure Legend Snippet:

    Techniques Used:

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    Alomone Labs anti-p2x7 receptor (extracellular)-fitc antibody
    Anti P2x7 Receptor (Extracellular) Fitc Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs anti p2x7 receptor extracellular fitc antibody
    A) Normalized MFI of HyLite Fluor 647-labeled Aβ1-42 peptide uptake normalized to DAPI in MDMi treated with DMSO alone (Ctrl), 1 mM ATP (ATP), 1 mM ATP and 10 uM A740003 (ATP + A74), or 10 uM A740003 (A74). Batch normalization was done in GraphPad PRISM 10 with 0% defined as the smallest mean in each data set and 100% as the average of all means in the data set. Statistics were determined with a repeated measure one-way ANOVA. N=26. B) Mean Intensity of HyLite Fluor 647-labeled Aβ1-42 per cell in MDMi imaged using confocal microscopy. MDMi were treated with DMSO alone (Ctrl), 1 mM ATP (ATP), or 1 mM ATP and 10 uM A740003 (ATP + A74). Statistics were determined with a repeated measure one-way ANOVA. N= 12. C) HyLite Fluor 647-labeled Aβ1-42 peptide uptake in MDMi treated with DMSO alone (Ctrl), or 500 uM bzATP (bzATP) as measured by fluorescence on a microplate reader. MFI= Mean Fluorescence Intensity normalized to DAPI. Batch normalization was done in GraphPad PRISM 10 with 0% defined as the smallest mean in each data set and 100% as the average of all means in the data set. Statistics were determined with a paired t-test. N=14. D) Representative images of MDMi treated with DMSO alone (Ctrl) or 1 mM ATP (ATP) and stained for <t>P2RX7</t> (green) and DAPI (blue) and used to measure uptake of HyLite Fluor 647-labeled Aβ1-42 peptide (red) and were processed in CellProfiler. ns= p>0.05, *p < 0.05, ** < 0.01,*** < 0.001, **** < 0.0001
    Anti P2x7 Receptor Extracellular Fitc Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs rabbit anti p2x7r monoclonal igg
    The sequences of oligonucleotide primers.
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    The sequences of oligonucleotide primers.
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    Alomone Labs antibodies anti p2x7 fitc
    <t>P2X7</t> receptors identified by immunofluorescence, in renal cortex and medulla in Sham and Ang II-infused rats. The expression of P2X7 receptors in the Ang II group is localized in tubular membranes ( A ). Western blot ( B ) showed abundance of the P2X7 receptor protein in the Ang II group ( n = 7 per group). * p < 0.01 vs. Sham. Ang II = angiotensin II, BBG = Brilliant blue G.
    Antibodies Anti P2x7 Fitc, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    <t>P2X7</t> receptors identified by immunofluorescence, in renal cortex and medulla in Sham and Ang II-infused rats. The expression of P2X7 receptors in the Ang II group is localized in tubular membranes ( A ). Western blot ( B ) showed abundance of the P2X7 receptor protein in the Ang II group ( n = 7 per group). * p < 0.01 vs. Sham. Ang II = angiotensin II, BBG = Brilliant blue G.
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    Alomone Labs anti p2x7r extracellular fitc
    P24XR and <t>P2X7R</t> Mediate ATP-Dependent Calcium Signal Propagation (A and B) Murine RAW 264.7 macrophages were loaded with photoactivable caged-IP 3 and the fluorescent calcium indicator Fluo-4 (green), and calcium signal propagation after IP 3 uncaging in the origin cell (white box) was monitored in live imaging. Experiments were performed in HBSS with 2 mM Ca 2+ (A) or in calcium-free HBSS supplemented with 2 mM EGTA (B). Scale bars, 50 μm. See also . (C and D) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 (C) or primary BMDMs (D). Error bars represent SEM. For statistical data analysis, Student’s t test was used ( ∗ p < 0.05). (E and F) Relative mRNA expression of different members of the P2X family of receptors, measured by real-time PCR in the RAW 264.7 cell line (E) and in BMDM primary macrophages (F). Error bars represent SEM. (G) Quantification of 5 independent live calcium imaging experiments with RAW 264.7 cells pre-treated with the P2X4R inhibitor 5BDBD (100 μM, 30 min at 37°C), the P2X7R inhibitor A740003 (100 μM, 30 min at 37°C), or their vehicle (DMSO) or left untreated. Error bars represent SEM. For statistical data analysis, One-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant). (H) Representative western blot (left) and quantification of repeated experiments (right) of P2X4R and P2X7R expression in RAW 264.7 cells transfected with siRNA specific for P2X4R and P2X7R or with scramble siRNA. Control cells were electroporated in the absence of oligonucleotides. (I) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 cells silenced for P2X4R and P2X7R. Error bars represent SEM. For statistical data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant).
    Anti P2x7r Extracellular Fitc, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    A) Normalized MFI of HyLite Fluor 647-labeled Aβ1-42 peptide uptake normalized to DAPI in MDMi treated with DMSO alone (Ctrl), 1 mM ATP (ATP), 1 mM ATP and 10 uM A740003 (ATP + A74), or 10 uM A740003 (A74). Batch normalization was done in GraphPad PRISM 10 with 0% defined as the smallest mean in each data set and 100% as the average of all means in the data set. Statistics were determined with a repeated measure one-way ANOVA. N=26. B) Mean Intensity of HyLite Fluor 647-labeled Aβ1-42 per cell in MDMi imaged using confocal microscopy. MDMi were treated with DMSO alone (Ctrl), 1 mM ATP (ATP), or 1 mM ATP and 10 uM A740003 (ATP + A74). Statistics were determined with a repeated measure one-way ANOVA. N= 12. C) HyLite Fluor 647-labeled Aβ1-42 peptide uptake in MDMi treated with DMSO alone (Ctrl), or 500 uM bzATP (bzATP) as measured by fluorescence on a microplate reader. MFI= Mean Fluorescence Intensity normalized to DAPI. Batch normalization was done in GraphPad PRISM 10 with 0% defined as the smallest mean in each data set and 100% as the average of all means in the data set. Statistics were determined with a paired t-test. N=14. D) Representative images of MDMi treated with DMSO alone (Ctrl) or 1 mM ATP (ATP) and stained for P2RX7 (green) and DAPI (blue) and used to measure uptake of HyLite Fluor 647-labeled Aβ1-42 peptide (red) and were processed in CellProfiler. ns= p>0.05, *p < 0.05, ** < 0.01,*** < 0.001, **** < 0.0001

    Journal: bioRxiv

    Article Title: IL-1RA Disrupts ATP Activation of P2RX7 in Human Monocyte-Derived Microglia-like Cells

    doi: 10.1101/2024.04.08.588607

    Figure Lengend Snippet: A) Normalized MFI of HyLite Fluor 647-labeled Aβ1-42 peptide uptake normalized to DAPI in MDMi treated with DMSO alone (Ctrl), 1 mM ATP (ATP), 1 mM ATP and 10 uM A740003 (ATP + A74), or 10 uM A740003 (A74). Batch normalization was done in GraphPad PRISM 10 with 0% defined as the smallest mean in each data set and 100% as the average of all means in the data set. Statistics were determined with a repeated measure one-way ANOVA. N=26. B) Mean Intensity of HyLite Fluor 647-labeled Aβ1-42 per cell in MDMi imaged using confocal microscopy. MDMi were treated with DMSO alone (Ctrl), 1 mM ATP (ATP), or 1 mM ATP and 10 uM A740003 (ATP + A74). Statistics were determined with a repeated measure one-way ANOVA. N= 12. C) HyLite Fluor 647-labeled Aβ1-42 peptide uptake in MDMi treated with DMSO alone (Ctrl), or 500 uM bzATP (bzATP) as measured by fluorescence on a microplate reader. MFI= Mean Fluorescence Intensity normalized to DAPI. Batch normalization was done in GraphPad PRISM 10 with 0% defined as the smallest mean in each data set and 100% as the average of all means in the data set. Statistics were determined with a paired t-test. N=14. D) Representative images of MDMi treated with DMSO alone (Ctrl) or 1 mM ATP (ATP) and stained for P2RX7 (green) and DAPI (blue) and used to measure uptake of HyLite Fluor 647-labeled Aβ1-42 peptide (red) and were processed in CellProfiler. ns= p>0.05, *p < 0.05, ** < 0.01,*** < 0.001, **** < 0.0001

    Article Snippet: PFA is removed and cells are PBS washed before staining with Anti-P2X7 Receptor (extracellular)-FITC antibody purchased from Alomone Labs (#APR-004).

    Techniques: Labeling, Confocal Microscopy, Fluorescence, Staining

    The sequences of oligonucleotide primers.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: The sequences of oligonucleotide primers.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques:

    The antibodies for immunoblotting.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: The antibodies for immunoblotting.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Western Blot, Concentration Assay

    The antibodies for fluorescence labeling.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: The antibodies for fluorescence labeling.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Fluorescence, Labeling, Concentration Assay

    Mechanical threshold and P2X7R and p-p38 expression. (a) Mechanical threshold after BTZ injection. (b, c) Western blot for P2X7R expression after BTZ treatment. (d) Immunofluorescence location of P2X7R in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. P2X7R is not expressed in NF-200-positive neurons. P2X7R is expressed in GFAP-labeled satellite glial cells (SGCs). (e) Immunofluorescence location of p-p38 in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. p-p38 is expressed in both MAP2-labeled neurons and GFAP-labeled SGCs. (f) Immunofluorescence location of P2X7R in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. P2X7R is expressed mainly in Iba-1-labeled microglial cells rather than in GFAP-labeled astrocytes and MAP2-labeled neurons. (g) Immunofluorescence location of p-p38 in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. p-p38 is expressed mainly in Iba-1-labeled microglial cells. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: Mechanical threshold and P2X7R and p-p38 expression. (a) Mechanical threshold after BTZ injection. (b, c) Western blot for P2X7R expression after BTZ treatment. (d) Immunofluorescence location of P2X7R in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. P2X7R is not expressed in NF-200-positive neurons. P2X7R is expressed in GFAP-labeled satellite glial cells (SGCs). (e) Immunofluorescence location of p-p38 in DRG. The arrows indicate the typical single- or double-labeled DRG neurons and satellite cells. p-p38 is expressed in both MAP2-labeled neurons and GFAP-labeled SGCs. (f) Immunofluorescence location of P2X7R in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. P2X7R is expressed mainly in Iba-1-labeled microglial cells rather than in GFAP-labeled astrocytes and MAP2-labeled neurons. (g) Immunofluorescence location of p-p38 in SDH. The arrows indicate the typical single-labeled and double-labeled cells in SDH. p-p38 is expressed mainly in Iba-1-labeled microglial cells. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Expressing, Injection, Western Blot, Immunofluorescence, Labeling

    p38 mRNA expression and p38 phosphorylation in DRG after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) p-p38 immunofluorescence labeling. The arrows show the typical p-p38 single-labeled DRG cells. (e) p-p38 fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗∗ P < 0.001.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: p38 mRNA expression and p38 phosphorylation in DRG after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) p-p38 immunofluorescence labeling. The arrows show the typical p-p38 single-labeled DRG cells. (e) p-p38 fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗∗ P < 0.001.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Expressing, Inhibition, Western Blot, Immunofluorescence, Labeling, Fluorescence

    p38 mRNA expression and p38 phosphorylation in SDH after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: p38 mRNA expression and p38 phosphorylation in SDH after inhibition of P2X7R with BBG. (a) p38 mRNA levels. (b) p-p38 protein immunoblotting bands. (c) p-p38 protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    IL-1 β , IL-6, and TNF- α mRNA expression in DRG and SDH after inhibition of P2X7R. (a) DRG IL-1 β mRNA. (b) DRG IL-6 mRNA. (c) DRG TNF- α mRNA. (d) SDH IL-1 β mRNA. (e) SDH IL-6 mRNA. (f) SDH TNF- α mRNA. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: IL-1 β , IL-6, and TNF- α mRNA expression in DRG and SDH after inhibition of P2X7R. (a) DRG IL-1 β mRNA. (b) DRG IL-6 mRNA. (c) DRG TNF- α mRNA. (d) SDH IL-1 β mRNA. (e) SDH IL-6 mRNA. (f) SDH TNF- α mRNA. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Expressing, Inhibition

    P2X7R mRNA and protein expression in DRG after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and GFAP coexpression fluorescence labeling for SGCs. The arrows indicate the typical single-labeled and double-labeled DRG satellite cells. (e) P2X7R and GFAP coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: P2X7R mRNA and protein expression in DRG after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and GFAP coexpression fluorescence labeling for SGCs. The arrows indicate the typical single-labeled and double-labeled DRG satellite cells. (e) P2X7R and GFAP coexpression fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    P2X7R mRNA and protein expression in SDH after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 colocalization fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: P2X7R mRNA and protein expression in SDH after inhibition of p38 phosphorylation. (a) P2X7R mRNA levels. (b) P2X7R protein immunoblotting bands. (c) P2X7R protein levels. (d) P2X7R and p-p38 coexpression fluorescence labeling. The arrows indicate the typical single-labeled and double-labeled SDH microglia. (e) P2X7R and p-p38 colocalization fluorescence density. Scale bar = 50 μ m. Mean ± SEM ( n = 5). ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001.

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Expressing, Inhibition, Western Blot, Fluorescence, Labeling

    Mechanical threshold alterations after inhibition of P2X7R or p38. (a) Mechanical threshold after inhibition of P2X7R. (b) Mechanical threshold after inhibition of p38. Mean ± SEM ( n = 5). ∗∗∗ P < 0.001 (vs. control); # P < 0.05; ## P < 0.01 (vs. BTZ group).

    Journal: BioMed Research International

    Article Title: The Actions and Mechanisms of P2X7R and p38 MAPK Activation in Mediating Bortezomib-Induced Neuropathic Pain

    doi: 10.1155/2020/8143754

    Figure Lengend Snippet: Mechanical threshold alterations after inhibition of P2X7R or p38. (a) Mechanical threshold after inhibition of P2X7R. (b) Mechanical threshold after inhibition of p38. Mean ± SEM ( n = 5). ∗∗∗ P < 0.001 (vs. control); # P < 0.05; ## P < 0.01 (vs. BTZ group).

    Article Snippet: Primary , Rabbit anti-P2X7R monoclonal IgG , 1 : 400 , Alomone Labs, Jerusalem, Israel.

    Techniques: Inhibition

    P2X7 receptors identified by immunofluorescence, in renal cortex and medulla in Sham and Ang II-infused rats. The expression of P2X7 receptors in the Ang II group is localized in tubular membranes ( A ). Western blot ( B ) showed abundance of the P2X7 receptor protein in the Ang II group ( n = 7 per group). * p < 0.01 vs. Sham. Ang II = angiotensin II, BBG = Brilliant blue G.

    Journal: International Journal of Molecular Sciences

    Article Title: The Role of P2X7 Purinergic Receptors in the Renal Inflammation Associated with Angiotensin II-Induced Hypertension

    doi: 10.3390/ijms21114041

    Figure Lengend Snippet: P2X7 receptors identified by immunofluorescence, in renal cortex and medulla in Sham and Ang II-infused rats. The expression of P2X7 receptors in the Ang II group is localized in tubular membranes ( A ). Western blot ( B ) showed abundance of the P2X7 receptor protein in the Ang II group ( n = 7 per group). * p < 0.01 vs. Sham. Ang II = angiotensin II, BBG = Brilliant blue G.

    Article Snippet: Kidney sections were incubated with 10% blocking serum in PBS for 1 h at room temperature followed by incubation at 4 °C overnight with the fluorescein isothiocyanate (FITC)-labeled antibodies anti-P2X7-FITC (Cat #: APR-008-F, rabbit polyclonal antibody, Alomone Laboratories, Jerusalem, Israel) at 1:250 [ ].

    Techniques: Immunofluorescence, Expressing, Western Blot

    P2X7 receptors (second column) in the infiltrating T cells of Ang II group co-localize with cluster differentiation (CD, first column) antigens in T cells (CD 3 positive cells), B cells (CD5 and CD20 positive cells), macrophages (CD68 positive cells) and leukocyte common antigen CD45, predominantly expressed in T lymphocytes.

    Journal: International Journal of Molecular Sciences

    Article Title: The Role of P2X7 Purinergic Receptors in the Renal Inflammation Associated with Angiotensin II-Induced Hypertension

    doi: 10.3390/ijms21114041

    Figure Lengend Snippet: P2X7 receptors (second column) in the infiltrating T cells of Ang II group co-localize with cluster differentiation (CD, first column) antigens in T cells (CD 3 positive cells), B cells (CD5 and CD20 positive cells), macrophages (CD68 positive cells) and leukocyte common antigen CD45, predominantly expressed in T lymphocytes.

    Article Snippet: Kidney sections were incubated with 10% blocking serum in PBS for 1 h at room temperature followed by incubation at 4 °C overnight with the fluorescein isothiocyanate (FITC)-labeled antibodies anti-P2X7-FITC (Cat #: APR-008-F, rabbit polyclonal antibody, Alomone Laboratories, Jerusalem, Israel) at 1:250 [ ].

    Techniques:

    P24XR and P2X7R Mediate ATP-Dependent Calcium Signal Propagation (A and B) Murine RAW 264.7 macrophages were loaded with photoactivable caged-IP 3 and the fluorescent calcium indicator Fluo-4 (green), and calcium signal propagation after IP 3 uncaging in the origin cell (white box) was monitored in live imaging. Experiments were performed in HBSS with 2 mM Ca 2+ (A) or in calcium-free HBSS supplemented with 2 mM EGTA (B). Scale bars, 50 μm. See also . (C and D) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 (C) or primary BMDMs (D). Error bars represent SEM. For statistical data analysis, Student’s t test was used ( ∗ p < 0.05). (E and F) Relative mRNA expression of different members of the P2X family of receptors, measured by real-time PCR in the RAW 264.7 cell line (E) and in BMDM primary macrophages (F). Error bars represent SEM. (G) Quantification of 5 independent live calcium imaging experiments with RAW 264.7 cells pre-treated with the P2X4R inhibitor 5BDBD (100 μM, 30 min at 37°C), the P2X7R inhibitor A740003 (100 μM, 30 min at 37°C), or their vehicle (DMSO) or left untreated. Error bars represent SEM. For statistical data analysis, One-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant). (H) Representative western blot (left) and quantification of repeated experiments (right) of P2X4R and P2X7R expression in RAW 264.7 cells transfected with siRNA specific for P2X4R and P2X7R or with scramble siRNA. Control cells were electroporated in the absence of oligonucleotides. (I) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 cells silenced for P2X4R and P2X7R. Error bars represent SEM. For statistical data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant).

    Journal: Cell Reports

    Article Title: Intercellular Calcium Signaling Induced by ATP Potentiates Macrophage Phagocytosis

    doi: 10.1016/j.celrep.2019.03.011

    Figure Lengend Snippet: P24XR and P2X7R Mediate ATP-Dependent Calcium Signal Propagation (A and B) Murine RAW 264.7 macrophages were loaded with photoactivable caged-IP 3 and the fluorescent calcium indicator Fluo-4 (green), and calcium signal propagation after IP 3 uncaging in the origin cell (white box) was monitored in live imaging. Experiments were performed in HBSS with 2 mM Ca 2+ (A) or in calcium-free HBSS supplemented with 2 mM EGTA (B). Scale bars, 50 μm. See also . (C and D) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 (C) or primary BMDMs (D). Error bars represent SEM. For statistical data analysis, Student’s t test was used ( ∗ p < 0.05). (E and F) Relative mRNA expression of different members of the P2X family of receptors, measured by real-time PCR in the RAW 264.7 cell line (E) and in BMDM primary macrophages (F). Error bars represent SEM. (G) Quantification of 5 independent live calcium imaging experiments with RAW 264.7 cells pre-treated with the P2X4R inhibitor 5BDBD (100 μM, 30 min at 37°C), the P2X7R inhibitor A740003 (100 μM, 30 min at 37°C), or their vehicle (DMSO) or left untreated. Error bars represent SEM. For statistical data analysis, One-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant). (H) Representative western blot (left) and quantification of repeated experiments (right) of P2X4R and P2X7R expression in RAW 264.7 cells transfected with siRNA specific for P2X4R and P2X7R or with scramble siRNA. Control cells were electroporated in the absence of oligonucleotides. (I) Quantification of 3 independent live calcium imaging experiments with RAW 264.7 cells silenced for P2X4R and P2X7R. Error bars represent SEM. For statistical data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant).

    Article Snippet: After counting, cells were incubated with anti-CD16/CD32 (BD PharMingen), and subsequently stained with the appropriate combinations of the following antibodies: anti-Cd11b-PerCP Cy5.5 (M1/70, BD Biosciences), anti-CD169-AlexaFluor647 (MOMA-1, Biorad), anti-P2X7R-extracellular-FITC (Alomone Labs), purified anti-P2X4R-extracellular (Alomone Labs) followed by incubation with the secondary anti-rabbit-FITC antibody (Thermo Fisher).

    Techniques: Imaging, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Transfection

    Macrophage Polarization Status Affects Calcium Signal Propagation (A) The surface expression of P2X4R (top) and P2X7R (bottom) was analyzed by flow cytometry in resting, IFNγ-treated (10 ng/mL, 24 h), or IL4-treated (20 ng/mL, 24 h) macrophages. Histograms show the quantification 3 independent biological replicates. Error bars represent SEM. For data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used (ns, non-significant; ∗∗ p < 0.01; ∗∗∗ p < 0.001). (B) Maximal back-projection of two representative live calcium imaging experiments performed with IFNγ- or IL4-treated RAW 264.7 cells loaded with caged-IP 3 and Fluo-4. The fluorescence variation 60 s after the irradiation of the origin cell (white box) is represented in false colors. (C) Representative traces of live calcium imaging experiments, showing the fluorescence variation after the uncaging of the origin cell (red) and the bystander macrophages (black). (D) Quantification of 3 independent live calcium imaging experiments. Error bars represent SEM. For data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant).

    Journal: Cell Reports

    Article Title: Intercellular Calcium Signaling Induced by ATP Potentiates Macrophage Phagocytosis

    doi: 10.1016/j.celrep.2019.03.011

    Figure Lengend Snippet: Macrophage Polarization Status Affects Calcium Signal Propagation (A) The surface expression of P2X4R (top) and P2X7R (bottom) was analyzed by flow cytometry in resting, IFNγ-treated (10 ng/mL, 24 h), or IL4-treated (20 ng/mL, 24 h) macrophages. Histograms show the quantification 3 independent biological replicates. Error bars represent SEM. For data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used (ns, non-significant; ∗∗ p < 0.01; ∗∗∗ p < 0.001). (B) Maximal back-projection of two representative live calcium imaging experiments performed with IFNγ- or IL4-treated RAW 264.7 cells loaded with caged-IP 3 and Fluo-4. The fluorescence variation 60 s after the irradiation of the origin cell (white box) is represented in false colors. (C) Representative traces of live calcium imaging experiments, showing the fluorescence variation after the uncaging of the origin cell (red) and the bystander macrophages (black). (D) Quantification of 3 independent live calcium imaging experiments. Error bars represent SEM. For data analysis, one-way ANOVA followed by Bonferroni’s multiple comparisons test was used ( ∗∗ p < 0.01; ns, non-significant).

    Article Snippet: After counting, cells were incubated with anti-CD16/CD32 (BD PharMingen), and subsequently stained with the appropriate combinations of the following antibodies: anti-Cd11b-PerCP Cy5.5 (M1/70, BD Biosciences), anti-CD169-AlexaFluor647 (MOMA-1, Biorad), anti-P2X7R-extracellular-FITC (Alomone Labs), purified anti-P2X4R-extracellular (Alomone Labs) followed by incubation with the secondary anti-rabbit-FITC antibody (Thermo Fisher).

    Techniques: Expressing, Flow Cytometry, Imaging, Fluorescence, Irradiation

    Extracellular ATP Is Required for Efficient Phagocytosis (A) Primary BMDMs were incubated with PhRodo E. coli fluorescent bioparticles in the presence or absence of 5 mM EGTA to chelate extracellular calcium. Phagocytosis was monitored at 15 or 30 min by flow cytometry (see <xref ref-type=Figure S4 ). Macrophages incubated with 20 μM cytochalasin D were used as negative reference. The phagocytic index was calculated as the percentage of fluorescent macrophages multiplied by their mean of fluorescence (MFI) and normalized on the cytochalasin-treated samples. (B) Primary BMDMs were loaded with the intracellular calcium chelator BAPTA-AM or its vehicle (loading solution) before performing the phagocytosis assay. (C) Primary BMDMs were incubated with PhRodo E. coli , PhRodo Zymosan, or PhRodo S. aureus fluorescent bioparticles, in the presence or absence of apyrase (5 U/mL). (D) Primary BMDMs were pretreated with the P2X4R inhibitor 5BDBD (100 μM), the P2X7R inhibitor A740003 (100 μM), or their vehicle (DMSO), or were left untreated, before performing the phagocytosis assay. (E) Phagocytosis was performed for 30 min in the presence or absence of MSC-derived EVs, pre-incubated or not with ARL-67516 (30 min, 200 μM). The graphs are representative of at least 3 independent biological replicates, each performed in technical triplicate. Error bars represent SEM. For data analysis, a two-way ANOVA followed by Tukey’s multiple comparisons test was used ( ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, non-significant). " width="100%" height="100%">

    Journal: Cell Reports

    Article Title: Intercellular Calcium Signaling Induced by ATP Potentiates Macrophage Phagocytosis

    doi: 10.1016/j.celrep.2019.03.011

    Figure Lengend Snippet: Extracellular ATP Is Required for Efficient Phagocytosis (A) Primary BMDMs were incubated with PhRodo E. coli fluorescent bioparticles in the presence or absence of 5 mM EGTA to chelate extracellular calcium. Phagocytosis was monitored at 15 or 30 min by flow cytometry (see Figure S4 ). Macrophages incubated with 20 μM cytochalasin D were used as negative reference. The phagocytic index was calculated as the percentage of fluorescent macrophages multiplied by their mean of fluorescence (MFI) and normalized on the cytochalasin-treated samples. (B) Primary BMDMs were loaded with the intracellular calcium chelator BAPTA-AM or its vehicle (loading solution) before performing the phagocytosis assay. (C) Primary BMDMs were incubated with PhRodo E. coli , PhRodo Zymosan, or PhRodo S. aureus fluorescent bioparticles, in the presence or absence of apyrase (5 U/mL). (D) Primary BMDMs were pretreated with the P2X4R inhibitor 5BDBD (100 μM), the P2X7R inhibitor A740003 (100 μM), or their vehicle (DMSO), or were left untreated, before performing the phagocytosis assay. (E) Phagocytosis was performed for 30 min in the presence or absence of MSC-derived EVs, pre-incubated or not with ARL-67516 (30 min, 200 μM). The graphs are representative of at least 3 independent biological replicates, each performed in technical triplicate. Error bars represent SEM. For data analysis, a two-way ANOVA followed by Tukey’s multiple comparisons test was used ( ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ns, non-significant).

    Article Snippet: After counting, cells were incubated with anti-CD16/CD32 (BD PharMingen), and subsequently stained with the appropriate combinations of the following antibodies: anti-Cd11b-PerCP Cy5.5 (M1/70, BD Biosciences), anti-CD169-AlexaFluor647 (MOMA-1, Biorad), anti-P2X7R-extracellular-FITC (Alomone Labs), purified anti-P2X4R-extracellular (Alomone Labs) followed by incubation with the secondary anti-rabbit-FITC antibody (Thermo Fisher).

    Techniques: Incubation, Flow Cytometry, Fluorescence, Phagocytosis Assay, Derivative Assay

    Journal: Cell Reports

    Article Title: Intercellular Calcium Signaling Induced by ATP Potentiates Macrophage Phagocytosis

    doi: 10.1016/j.celrep.2019.03.011

    Figure Lengend Snippet:

    Article Snippet: After counting, cells were incubated with anti-CD16/CD32 (BD PharMingen), and subsequently stained with the appropriate combinations of the following antibodies: anti-Cd11b-PerCP Cy5.5 (M1/70, BD Biosciences), anti-CD169-AlexaFluor647 (MOMA-1, Biorad), anti-P2X7R-extracellular-FITC (Alomone Labs), purified anti-P2X4R-extracellular (Alomone Labs) followed by incubation with the secondary anti-rabbit-FITC antibody (Thermo Fisher).

    Techniques: Recombinant, Negative Control, Real-time Polymerase Chain Reaction, Software

    Journal: Cell Reports

    Article Title: Intercellular Calcium Signaling Induced by ATP Potentiates Macrophage Phagocytosis

    doi: 10.1016/j.celrep.2019.03.011

    Figure Lengend Snippet:

    Article Snippet: After counting, cells were incubated with anti-CD16/CD32 (BD PharMingen), and subsequently stained with the appropriate combinations of the following antibodies: anti-Cd11b-PerCP Cy5.5 (M1/70, BD Biosciences), anti-CD169-AlexaFluor647 (MOMA-1, Biorad), anti-P2X7R-extracellular-FITC (Alomone Labs), purified anti-P2X4R-extracellular (Alomone Labs) followed by incubation with the secondary anti-rabbit-FITC antibody (Thermo Fisher).

    Techniques: