goat f  (SouthernBiotech)

 
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  • 93
    Name:
    Goat F ab 2 IgG PE
    Description:

    Catalog Number:
    0110-09
    Price:
    None
    Applications:
    Quality tested applications for relevant formats include -Flow Cytometry 1-3ELISAFLISAOther referenced applications for relevant formats include -Immunocytochemistry 4
    Source:
    Pepsin digest of Goat IgG (SB Cat. No. 0109)
    Format:
    PE (R-phycoerythrin)
    Buy from Supplier


    Structured Review

    SouthernBiotech goat f
    Goat F ab 2 IgG PE

    https://www.bioz.com/result/goat f/product/SouthernBiotech
    Average 93 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    goat f - by Bioz Stars, 2020-09
    93/100 stars

    Images

    Related Articles

    Transduction:

    Article Title: Coexpressed Catalase Protects Chimeric Antigen Receptor–Redirected T Cells as well as Bystander Cells from Oxidative Stress–Induced Loss of Antitumor Activity
    Article Snippet: .. Activated lymphocytes were cocultured with virus producing transiently transfected HEK293T cells for 48 h. Transduction efficiency was analyzed by flow cytometry with goat F(ab′)2 anti-human IgG-PE (SouthernBiotech) and mouse anti-human CD3-allophycocyanin (BioLegend). .. Transduced T cells were sorted using anti-PE MACS (Miltenyi Biotec) beads in combination with F(ab′)2 anti-human IgG-PE (Southern Biotech).

    Transfection:

    Article Title: Coexpressed Catalase Protects Chimeric Antigen Receptor–Redirected T Cells as well as Bystander Cells from Oxidative Stress–Induced Loss of Antitumor Activity
    Article Snippet: .. Activated lymphocytes were cocultured with virus producing transiently transfected HEK293T cells for 48 h. Transduction efficiency was analyzed by flow cytometry with goat F(ab′)2 anti-human IgG-PE (SouthernBiotech) and mouse anti-human CD3-allophycocyanin (BioLegend). .. Transduced T cells were sorted using anti-PE MACS (Miltenyi Biotec) beads in combination with F(ab′)2 anti-human IgG-PE (Southern Biotech).

    Fluorescence:

    Article Title: Expression of ORAI1, a Plasma Membrane Resident Subunit of the CRAC Channel, in Rodent and Non-rodent Species
    Article Snippet: .. Monoclonal antibodies were partially purified from the expanded hybridoma cultures, and specific binding was confirmed through FMAT and FACS (fluorescence-activated cell sorting), where cross-species reactivity was also evaluated using methods previously described , using goat F(ab’)2 anti-mouse IgG-phycoerythrin (IgG-PE) as the detection reagent (Southern Biotech; Birmingham, AL; Cat# 1030-009). .. After single-cell sorting and specificity characterization, the hybridoma line 266 was identified as a promising candidate as it did not bind to either ORAI2 or ORAI3 using FACS with cell lines expressing those family members.

    Cytometry:

    Article Title: Coexpressed Catalase Protects Chimeric Antigen Receptor–Redirected T Cells as well as Bystander Cells from Oxidative Stress–Induced Loss of Antitumor Activity
    Article Snippet: .. Activated lymphocytes were cocultured with virus producing transiently transfected HEK293T cells for 48 h. Transduction efficiency was analyzed by flow cytometry with goat F(ab′)2 anti-human IgG-PE (SouthernBiotech) and mouse anti-human CD3-allophycocyanin (BioLegend). .. Transduced T cells were sorted using anti-PE MACS (Miltenyi Biotec) beads in combination with F(ab′)2 anti-human IgG-PE (Southern Biotech).

    Purification:

    Article Title: Expression of ORAI1, a Plasma Membrane Resident Subunit of the CRAC Channel, in Rodent and Non-rodent Species
    Article Snippet: .. Monoclonal antibodies were partially purified from the expanded hybridoma cultures, and specific binding was confirmed through FMAT and FACS (fluorescence-activated cell sorting), where cross-species reactivity was also evaluated using methods previously described , using goat F(ab’)2 anti-mouse IgG-phycoerythrin (IgG-PE) as the detection reagent (Southern Biotech; Birmingham, AL; Cat# 1030-009). .. After single-cell sorting and specificity characterization, the hybridoma line 266 was identified as a promising candidate as it did not bind to either ORAI2 or ORAI3 using FACS with cell lines expressing those family members.

    other:

    Article Title: CLEC9A modulates macrophage-mediated neutrophil recruitment in response to heat-killed Mycobacterium tuberculosis H37Ra
    Article Snippet: Goat F(ab')2 Anti-Human IgG-PE (2042–09) was obtained from Southern Biotech.

    FACS:

    Article Title: Expression of ORAI1, a Plasma Membrane Resident Subunit of the CRAC Channel, in Rodent and Non-rodent Species
    Article Snippet: .. Monoclonal antibodies were partially purified from the expanded hybridoma cultures, and specific binding was confirmed through FMAT and FACS (fluorescence-activated cell sorting), where cross-species reactivity was also evaluated using methods previously described , using goat F(ab’)2 anti-mouse IgG-phycoerythrin (IgG-PE) as the detection reagent (Southern Biotech; Birmingham, AL; Cat# 1030-009). .. After single-cell sorting and specificity characterization, the hybridoma line 266 was identified as a promising candidate as it did not bind to either ORAI2 or ORAI3 using FACS with cell lines expressing those family members.

    Binding Assay:

    Article Title: Expression of ORAI1, a Plasma Membrane Resident Subunit of the CRAC Channel, in Rodent and Non-rodent Species
    Article Snippet: .. Monoclonal antibodies were partially purified from the expanded hybridoma cultures, and specific binding was confirmed through FMAT and FACS (fluorescence-activated cell sorting), where cross-species reactivity was also evaluated using methods previously described , using goat F(ab’)2 anti-mouse IgG-phycoerythrin (IgG-PE) as the detection reagent (Southern Biotech; Birmingham, AL; Cat# 1030-009). .. After single-cell sorting and specificity characterization, the hybridoma line 266 was identified as a promising candidate as it did not bind to either ORAI2 or ORAI3 using FACS with cell lines expressing those family members.

    Flow Cytometry:

    Article Title: Coexpressed Catalase Protects Chimeric Antigen Receptor–Redirected T Cells as well as Bystander Cells from Oxidative Stress–Induced Loss of Antitumor Activity
    Article Snippet: .. Activated lymphocytes were cocultured with virus producing transiently transfected HEK293T cells for 48 h. Transduction efficiency was analyzed by flow cytometry with goat F(ab′)2 anti-human IgG-PE (SouthernBiotech) and mouse anti-human CD3-allophycocyanin (BioLegend). .. Transduced T cells were sorted using anti-PE MACS (Miltenyi Biotec) beads in combination with F(ab′)2 anti-human IgG-PE (Southern Biotech).

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  • 84
    SouthernBiotech dylight 650 conjugated goat f ab 2 anti human λ κ
    Atypical MBCs rapidly capture and internalize PMS-bound <t>anti-λ/κ.</t> PBMCs were incubated for increasing lengths of time (30 to 120 min) at 37°C on PMS containing biotinylated anti-λ/κ conjugated to <t>DyLight</t> 650 and avidin-pHrodo. At each time point, cells were harvested and stained with streptavidin-Alexa488 and antibodies specific for CD19, CD21, and CD27 to identify naïve B cells (CD19 + CD21 + CD27 − ), classical MBCs (CD19 + CD21 + CD27 + ), and atypical MBCs (CD19 + CD21 − CD27 − ). Antigen capture, internalization, and trafficking into the acidic compartments were analyzed by flow cytometry as described in the text and detailed in Materials and Methods ( n = 4). In all cases: atypical MBCs (red circles), classical MBCs (green circles), and naïve B cells (blue circles) (see also fig. S2). ( A ) Comparison of the total amount of anti-λ/κ captured from PMS over time indicated by the MFI of DyLight 650. ( B ) Comparison of the amount of anti-λ/κ internalized in 15 min (left) and over time (right). ( C ) Comparison of anti-λ/κ trafficking to acidic compartment over time indicated by the percentage of cells positive for pHrodo or by the increases in pHrodo MFI from time 0. Data were analyzed using paired t test (A and C) or two-way analysis of variance (ANOVA) and Tukey’s multiple comparison test (B). * P
    Dylight 650 Conjugated Goat F Ab 2 Anti Human λ κ, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dylight 650 conjugated goat f ab 2 anti human λ κ/product/SouthernBiotech
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dylight 650 conjugated goat f ab 2 anti human λ κ - by Bioz Stars, 2020-09
    84/100 stars
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    94
    SouthernBiotech goat f ab 2 anti human iga alexa fluor 647
    Binding of type 3 PV to OCMS hybridomas secreting anti-PV <t>IgA</t> mAbs. Hybridomas were incubated overnight with the rabbit mAb specific for human IgA and then with biotinylated PV. Human IgA and PV3 binding to the cells were detected with an <t>Alexa-Fluor</t> 647 anti-human IgA secondary and Alexa-Fluor 488 conjugated streptavidin, respectively. Nuclei were stained with DAPI. Scale bar = 10 µm.
    Goat F Ab 2 Anti Human Iga Alexa Fluor 647, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat f ab 2 anti human iga alexa fluor 647/product/SouthernBiotech
    Average 94 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    goat f ab 2 anti human iga alexa fluor 647 - by Bioz Stars, 2020-09
    94/100 stars
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    84
    SouthernBiotech alexa fluor647 conjugated goat f ab 2 anti human kappa
    The binding of SpA-B to antigen-bound IgG decreases C1q binding on target surfaces even if C1q is incubated before SpA-B. A Binding of anti-DNP IgG1-WT and IgG3-WT to DNP-coated beads, detected with <t>Alexa</t> <t>Fluor647-conjugated</t> goat F(ab’) 2 anti-human kappa by flow-cytometry. B Binding of SpA-B to anti-DNP IgG1-WT and IgG3-WT bound to DNP-coated beads, detected with anti-HexaHistidine chicken antibody by flow cytometry. C, D C1q binding on IgG1-WT and IgG3-WT bound to DNP-coated beads, detected with FITC-conjugated rabbit F(ab’) 2 anti-human C1q by flow-cytometry. Buffer (solid lines) or SpA-B (dotted lines) was added to the beads only after C1q (C) or C1 complex (D). Bars represent the same data for the 20 nM IgG concentration only and the black dotted line shows the background fluorescence from beads that were not incubated with IgG. Data are presented as geometric means ± SD of three (C) or four (D) independent experiments. Statistical analysis was performed using an unpaired two-tailed t-test to compare buffer and SpA-B conditions and showed when significant as **P ≤ 0.01, ***P≤ 0.001 and ****P ≤ 0.0001.
    Alexa Fluor647 Conjugated Goat F Ab 2 Anti Human Kappa, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/alexa fluor647 conjugated goat f ab 2 anti human kappa/product/SouthernBiotech
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    alexa fluor647 conjugated goat f ab 2 anti human kappa - by Bioz Stars, 2020-09
    84/100 stars
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    97
    SouthernBiotech goat anti human igm
    Internalization of BCR changes downstream signaling events upon restimulation. EU12 μ HC + , Daudi, and Ramos cells were directly stimulated with <t>Cy5</t> labeled <t>anti-IgM</t> antibodies (10 μ g/mL) for 3 min or pretreated with goat anti-IgM antibody F(ab′) 2 fragments (2 μ g/mL) for 30 min, washed, recovered for 4 h, and restimulated with Cy5 labeled anti-IgM antibodies (10 μ g/mL) for 3 min. Fixed and permeabilized cells were analyzed by immunofluorescence microscopy for phosphorylated SYK (a) and ERK1/2 (b). Scale bar, 10 μ m. Results shown are representatives from three independent experiments with more than 150 cells analyzed.
    Goat Anti Human Igm, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 97/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti human igm/product/SouthernBiotech
    Average 97 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    goat anti human igm - by Bioz Stars, 2020-09
    97/100 stars
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    Image Search Results


    Atypical MBCs rapidly capture and internalize PMS-bound anti-λ/κ. PBMCs were incubated for increasing lengths of time (30 to 120 min) at 37°C on PMS containing biotinylated anti-λ/κ conjugated to DyLight 650 and avidin-pHrodo. At each time point, cells were harvested and stained with streptavidin-Alexa488 and antibodies specific for CD19, CD21, and CD27 to identify naïve B cells (CD19 + CD21 + CD27 − ), classical MBCs (CD19 + CD21 + CD27 + ), and atypical MBCs (CD19 + CD21 − CD27 − ). Antigen capture, internalization, and trafficking into the acidic compartments were analyzed by flow cytometry as described in the text and detailed in Materials and Methods ( n = 4). In all cases: atypical MBCs (red circles), classical MBCs (green circles), and naïve B cells (blue circles) (see also fig. S2). ( A ) Comparison of the total amount of anti-λ/κ captured from PMS over time indicated by the MFI of DyLight 650. ( B ) Comparison of the amount of anti-λ/κ internalized in 15 min (left) and over time (right). ( C ) Comparison of anti-λ/κ trafficking to acidic compartment over time indicated by the percentage of cells positive for pHrodo or by the increases in pHrodo MFI from time 0. Data were analyzed using paired t test (A and C) or two-way analysis of variance (ANOVA) and Tukey’s multiple comparison test (B). * P

    Journal: Science Advances

    Article Title: Expression of inhibitory receptors by B cells in chronic human infectious diseases restricts responses to membrane-associated antigens

    doi: 10.1126/sciadv.aba6493

    Figure Lengend Snippet: Atypical MBCs rapidly capture and internalize PMS-bound anti-λ/κ. PBMCs were incubated for increasing lengths of time (30 to 120 min) at 37°C on PMS containing biotinylated anti-λ/κ conjugated to DyLight 650 and avidin-pHrodo. At each time point, cells were harvested and stained with streptavidin-Alexa488 and antibodies specific for CD19, CD21, and CD27 to identify naïve B cells (CD19 + CD21 + CD27 − ), classical MBCs (CD19 + CD21 + CD27 + ), and atypical MBCs (CD19 + CD21 − CD27 − ). Antigen capture, internalization, and trafficking into the acidic compartments were analyzed by flow cytometry as described in the text and detailed in Materials and Methods ( n = 4). In all cases: atypical MBCs (red circles), classical MBCs (green circles), and naïve B cells (blue circles) (see also fig. S2). ( A ) Comparison of the total amount of anti-λ/κ captured from PMS over time indicated by the MFI of DyLight 650. ( B ) Comparison of the amount of anti-λ/κ internalized in 15 min (left) and over time (right). ( C ) Comparison of anti-λ/κ trafficking to acidic compartment over time indicated by the percentage of cells positive for pHrodo or by the increases in pHrodo MFI from time 0. Data were analyzed using paired t test (A and C) or two-way analysis of variance (ANOVA) and Tukey’s multiple comparison test (B). * P

    Article Snippet: After washing with 10 ml of 1× PMS washing buffer to remove all debris and incubating with the blocking buffer (5% BSA-containing 1× PMS buffer) for 30 min, annexin V–biotin (0.5 mg/ml) was added for binding to the PMS for 30 min and washed with 15 ml of 1× washing buffer, and this step was repeated with streptavidin (5 mg/ml) and then 10 nM biotin- and DyLight 650–conjugated goat F(ab′)2 anti-human λ + κ (SouthernBiotech).

    Techniques: Incubation, Avidin-Biotin Assay, Staining, Flow Cytometry

    Binding of type 3 PV to OCMS hybridomas secreting anti-PV IgA mAbs. Hybridomas were incubated overnight with the rabbit mAb specific for human IgA and then with biotinylated PV. Human IgA and PV3 binding to the cells were detected with an Alexa-Fluor 647 anti-human IgA secondary and Alexa-Fluor 488 conjugated streptavidin, respectively. Nuclei were stained with DAPI. Scale bar = 10 µm.

    Journal: Antibodies

    Article Title: Human IgA Monoclonal Antibodies That Neutralize Poliovirus, Produced by Hybridomas and Recombinant Expression

    doi: 10.3390/antib9010005

    Figure Lengend Snippet: Binding of type 3 PV to OCMS hybridomas secreting anti-PV IgA mAbs. Hybridomas were incubated overnight with the rabbit mAb specific for human IgA and then with biotinylated PV. Human IgA and PV3 binding to the cells were detected with an Alexa-Fluor 647 anti-human IgA secondary and Alexa-Fluor 488 conjugated streptavidin, respectively. Nuclei were stained with DAPI. Scale bar = 10 µm.

    Article Snippet: The cells were washed 3 times with PBS 1% BSA, co-incubated for 1 h with 1:200 Alexa Fluor 647 conjugated goat F(ab’)2 anti-human IgA (2052-31; SouthernBiotech) and 1:200 Alexa Fluor 488 conjugated AffiniPure F(ab’)2 fragment goat anti-rabbit IgG (H+L) (111-546-144; Jackson ImmunoResearch) to detect the expression of IgA and OCMS respectively.

    Techniques: Binding Assay, Incubation, Staining

    Expression of recombinant IgA in 293T OCMS cells. 293T OCMS ( top row ) and 293T ( bottom row ) cells were transiently transfected with plasmids encoding recombinant 1G1 IgA HC and LC genes. The next day, the cells were incubated overnight with the rabbit anti-human IgA mAb (Linker). Bound Linker was detected with Alexa-Fluor 488 anti-rabbit IgG secondary antibody (green). Captured human IgA was labeled with Alexa-Fluor 647 anti human IgA (magenta). Nuclei were stained with DAPI (blue). Scale bar = 5 µm.

    Journal: Antibodies

    Article Title: Human IgA Monoclonal Antibodies That Neutralize Poliovirus, Produced by Hybridomas and Recombinant Expression

    doi: 10.3390/antib9010005

    Figure Lengend Snippet: Expression of recombinant IgA in 293T OCMS cells. 293T OCMS ( top row ) and 293T ( bottom row ) cells were transiently transfected with plasmids encoding recombinant 1G1 IgA HC and LC genes. The next day, the cells were incubated overnight with the rabbit anti-human IgA mAb (Linker). Bound Linker was detected with Alexa-Fluor 488 anti-rabbit IgG secondary antibody (green). Captured human IgA was labeled with Alexa-Fluor 647 anti human IgA (magenta). Nuclei were stained with DAPI (blue). Scale bar = 5 µm.

    Article Snippet: The cells were washed 3 times with PBS 1% BSA, co-incubated for 1 h with 1:200 Alexa Fluor 647 conjugated goat F(ab’)2 anti-human IgA (2052-31; SouthernBiotech) and 1:200 Alexa Fluor 488 conjugated AffiniPure F(ab’)2 fragment goat anti-rabbit IgG (H+L) (111-546-144; Jackson ImmunoResearch) to detect the expression of IgA and OCMS respectively.

    Techniques: Expressing, Recombinant, Transfection, Incubation, Labeling, Staining

    The binding of SpA-B to antigen-bound IgG decreases C1q binding on target surfaces even if C1q is incubated before SpA-B. A Binding of anti-DNP IgG1-WT and IgG3-WT to DNP-coated beads, detected with Alexa Fluor647-conjugated goat F(ab’) 2 anti-human kappa by flow-cytometry. B Binding of SpA-B to anti-DNP IgG1-WT and IgG3-WT bound to DNP-coated beads, detected with anti-HexaHistidine chicken antibody by flow cytometry. C, D C1q binding on IgG1-WT and IgG3-WT bound to DNP-coated beads, detected with FITC-conjugated rabbit F(ab’) 2 anti-human C1q by flow-cytometry. Buffer (solid lines) or SpA-B (dotted lines) was added to the beads only after C1q (C) or C1 complex (D). Bars represent the same data for the 20 nM IgG concentration only and the black dotted line shows the background fluorescence from beads that were not incubated with IgG. Data are presented as geometric means ± SD of three (C) or four (D) independent experiments. Statistical analysis was performed using an unpaired two-tailed t-test to compare buffer and SpA-B conditions and showed when significant as **P ≤ 0.01, ***P≤ 0.001 and ****P ≤ 0.0001.

    Journal: bioRxiv

    Article Title: Staphylococcal protein A inhibits complement activation by interfering with IgG hexamer formation

    doi: 10.1101/2020.07.20.212118

    Figure Lengend Snippet: The binding of SpA-B to antigen-bound IgG decreases C1q binding on target surfaces even if C1q is incubated before SpA-B. A Binding of anti-DNP IgG1-WT and IgG3-WT to DNP-coated beads, detected with Alexa Fluor647-conjugated goat F(ab’) 2 anti-human kappa by flow-cytometry. B Binding of SpA-B to anti-DNP IgG1-WT and IgG3-WT bound to DNP-coated beads, detected with anti-HexaHistidine chicken antibody by flow cytometry. C, D C1q binding on IgG1-WT and IgG3-WT bound to DNP-coated beads, detected with FITC-conjugated rabbit F(ab’) 2 anti-human C1q by flow-cytometry. Buffer (solid lines) or SpA-B (dotted lines) was added to the beads only after C1q (C) or C1 complex (D). Bars represent the same data for the 20 nM IgG concentration only and the black dotted line shows the background fluorescence from beads that were not incubated with IgG. Data are presented as geometric means ± SD of three (C) or four (D) independent experiments. Statistical analysis was performed using an unpaired two-tailed t-test to compare buffer and SpA-B conditions and showed when significant as **P ≤ 0.01, ***P≤ 0.001 and ****P ≤ 0.0001.

    Article Snippet: For antibody binding detection, IgG-bound DNP-beads were next incubated with 1 µg/mL Alexa Fluor647 -conjugated goat F(ab’)2 anti-human kappa (Southern Biotech) in PBS-TH.

    Techniques: Binding Assay, Incubation, Flow Cytometry, Concentration Assay, Fluorescence, Two Tailed Test

    Internalization of BCR changes downstream signaling events upon restimulation. EU12 μ HC + , Daudi, and Ramos cells were directly stimulated with Cy5 labeled anti-IgM antibodies (10 μ g/mL) for 3 min or pretreated with goat anti-IgM antibody F(ab′) 2 fragments (2 μ g/mL) for 30 min, washed, recovered for 4 h, and restimulated with Cy5 labeled anti-IgM antibodies (10 μ g/mL) for 3 min. Fixed and permeabilized cells were analyzed by immunofluorescence microscopy for phosphorylated SYK (a) and ERK1/2 (b). Scale bar, 10 μ m. Results shown are representatives from three independent experiments with more than 150 cells analyzed.

    Journal: BioMed Research International

    Article Title: Internalization of B Cell Receptors in Human EU12 μHC+ Immature B Cells Specifically Alters Downstream Signaling Events

    doi: 10.1155/2013/807240

    Figure Lengend Snippet: Internalization of BCR changes downstream signaling events upon restimulation. EU12 μ HC + , Daudi, and Ramos cells were directly stimulated with Cy5 labeled anti-IgM antibodies (10 μ g/mL) for 3 min or pretreated with goat anti-IgM antibody F(ab′) 2 fragments (2 μ g/mL) for 30 min, washed, recovered for 4 h, and restimulated with Cy5 labeled anti-IgM antibodies (10 μ g/mL) for 3 min. Fixed and permeabilized cells were analyzed by immunofluorescence microscopy for phosphorylated SYK (a) and ERK1/2 (b). Scale bar, 10 μ m. Results shown are representatives from three independent experiments with more than 150 cells analyzed.

    Article Snippet: Antibodies and Reagents Immunofluorescence assays were performed using the following antibodies: Cy5-labeled AffiniPure donkey anti-human IgM F(ab′)2 fragment (Jackson ImmunoResearch, West Grove, PA), goat anti-human IgM (μ chain specific, LE/AF) F(ab′)2 fragments (Southern Biotech, Birmingham, AL), goat anti-rabbit Alexa-488 (Invitrogen, Carlsbad, CA), anti-phospho-SYK, phospho-ERK, and phospho-CD19 antibodies (Cell Signaling Technology, Danvers, MA).

    Techniques: Labeling, Immunofluorescence, Microscopy

    Anti-IgM antibody induces BCR internalization in EU12 μ HC + immature B cells and in mature Daudi or Ramos B cells. (a) Human U12 μ HC + , Daudi, and Ramos cells were treated with goat anti-IgM antibody F(ab′) 2 fragments (2 μ g/mL) for different times. The cells were stained with the Cy5 labeled anti-IgM antibodies at different time points and subjected to flow cytometric analysis. The percentages of cells expressing cell surface IgM are indicated. (b) Murine 70Z/3 cells were treated with goat anti-IgM antibodies (5 μ g/mL) for different times; cell surface IgM were monitored by APC labeled anti-IgM antibodies. The percentages of cells expressing cell surface IgM are indicated. Unstained samples were used as negative control. (c) Immunofluorescence analysis. For untreated cells, EU12 μ HC + , Daudi, or Ramos cells were fixed and stained with Cy5 labeled anti-IgM antibodies. For anti-IgM antibody treated cells, EU12 μ HC + , Daudi, and Ramos cells were treated with Cy5 labeled anti-IgM (2 μ g/mL) for 30 min. Cells were then fixed and visualized on glass slides with fluorescence microscopy. (d) Human EU12 μ HC + or Daudi cells were treated with anti-IgM antibodies (2 μ g/mL) for 24 hours and cell surface HLA and CD86 levels were analyzed by flow cytometry.

    Journal: BioMed Research International

    Article Title: Internalization of B Cell Receptors in Human EU12 μHC+ Immature B Cells Specifically Alters Downstream Signaling Events

    doi: 10.1155/2013/807240

    Figure Lengend Snippet: Anti-IgM antibody induces BCR internalization in EU12 μ HC + immature B cells and in mature Daudi or Ramos B cells. (a) Human U12 μ HC + , Daudi, and Ramos cells were treated with goat anti-IgM antibody F(ab′) 2 fragments (2 μ g/mL) for different times. The cells were stained with the Cy5 labeled anti-IgM antibodies at different time points and subjected to flow cytometric analysis. The percentages of cells expressing cell surface IgM are indicated. (b) Murine 70Z/3 cells were treated with goat anti-IgM antibodies (5 μ g/mL) for different times; cell surface IgM were monitored by APC labeled anti-IgM antibodies. The percentages of cells expressing cell surface IgM are indicated. Unstained samples were used as negative control. (c) Immunofluorescence analysis. For untreated cells, EU12 μ HC + , Daudi, or Ramos cells were fixed and stained with Cy5 labeled anti-IgM antibodies. For anti-IgM antibody treated cells, EU12 μ HC + , Daudi, and Ramos cells were treated with Cy5 labeled anti-IgM (2 μ g/mL) for 30 min. Cells were then fixed and visualized on glass slides with fluorescence microscopy. (d) Human EU12 μ HC + or Daudi cells were treated with anti-IgM antibodies (2 μ g/mL) for 24 hours and cell surface HLA and CD86 levels were analyzed by flow cytometry.

    Article Snippet: Antibodies and Reagents Immunofluorescence assays were performed using the following antibodies: Cy5-labeled AffiniPure donkey anti-human IgM F(ab′)2 fragment (Jackson ImmunoResearch, West Grove, PA), goat anti-human IgM (μ chain specific, LE/AF) F(ab′)2 fragments (Southern Biotech, Birmingham, AL), goat anti-rabbit Alexa-488 (Invitrogen, Carlsbad, CA), anti-phospho-SYK, phospho-ERK, and phospho-CD19 antibodies (Cell Signaling Technology, Danvers, MA).

    Techniques: Staining, Labeling, Flow Cytometry, Expressing, Negative Control, Immunofluorescence, Fluorescence, Microscopy, Cytometry