anti human cxcr4  (Santa Cruz Biotechnology)

 
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    Name:
    CXCR 4 Antibody
    Description:
    Anti CXCR 4 Antibody 4G10 is a mouse monoclonal IgG1 kappa light chain CXCR 4 antibody provided at 200 µg ml raised against the N terminus amino acids of CXCR 4 of human origin Anti CXCR 4 Antibody 4G10 is recommended for detection of CXCR 4 of mouse rat and human origin by WB IP IF IHC P and FCM Anti CXCR 4 Antibody 4G10 is available conjugated to agarose for IP HRP for WB IHC P and ELISA and to either phycoerythrin or FITC for IF IHC P and FCM also available conjugated to Alexa Fluor 488 Alexa Fluor 546 Alexa Fluor 594 or Alexa Fluor 647 for WB RGB IF IHC P and FCM and for use with RGB fluorescent imaging systems such as iBright FL1000 FluorChem Typhoon Azure and other comparable systems also available conjugated to Alexa Fluor 680 or Alexa Fluor 790 for WB NIR IF and FCM for use with Near Infrared NIR detection systems such as LI COR Odyssey iBright FL1000 FluorChem Typhoon Azure and other comparable systems Contact our Technical Service Department or your local Distributor for more information on how to receive a FREE 10 µg sample of CXCR 4 4G10 sc 53534
    Catalog Number:
    SC-53534
    Price:
    None
    Category:
    Antibodies Primary Antibodies and ImmunoCruz Conjugates Membrane Receptors CXCR Antibodies CXCR 4 Antibody 4G10
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    Structured Review

    Santa Cruz Biotechnology anti human cxcr4
    Not unlike the NST (see ), virtually all neurons in the AP express <t>CXCR4</t> receptors and astrocytes do not. (A) Cholinergic (ChAT-ir; red) AP neurons express CXCR4 receptors (green); colocalization appears yellow (examples at arrows). (B) Catecholaminergic
    Anti CXCR 4 Antibody 4G10 is a mouse monoclonal IgG1 kappa light chain CXCR 4 antibody provided at 200 µg ml raised against the N terminus amino acids of CXCR 4 of human origin Anti CXCR 4 Antibody 4G10 is recommended for detection of CXCR 4 of mouse rat and human origin by WB IP IF IHC P and FCM Anti CXCR 4 Antibody 4G10 is available conjugated to agarose for IP HRP for WB IHC P and ELISA and to either phycoerythrin or FITC for IF IHC P and FCM also available conjugated to Alexa Fluor 488 Alexa Fluor 546 Alexa Fluor 594 or Alexa Fluor 647 for WB RGB IF IHC P and FCM and for use with RGB fluorescent imaging systems such as iBright FL1000 FluorChem Typhoon Azure and other comparable systems also available conjugated to Alexa Fluor 680 or Alexa Fluor 790 for WB NIR IF and FCM for use with Near Infrared NIR detection systems such as LI COR Odyssey iBright FL1000 FluorChem Typhoon Azure and other comparable systems Contact our Technical Service Department or your local Distributor for more information on how to receive a FREE 10 µg sample of CXCR 4 4G10 sc 53534
    https://www.bioz.com/result/anti human cxcr4/product/Santa Cruz Biotechnology
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti human cxcr4 - by Bioz Stars, 2021-09
    95/100 stars

    Images

    1) Product Images from "CXCR4 receptors in the dorsal medulla: implications for autonomic dysfunction"

    Article Title: CXCR4 receptors in the dorsal medulla: implications for autonomic dysfunction

    Journal:

    doi: 10.1111/j.1460-9568.2008.06058.x

    Not unlike the NST (see ), virtually all neurons in the AP express CXCR4 receptors and astrocytes do not. (A) Cholinergic (ChAT-ir; red) AP neurons express CXCR4 receptors (green); colocalization appears yellow (examples at arrows). (B) Catecholaminergic
    Figure Legend Snippet: Not unlike the NST (see ), virtually all neurons in the AP express CXCR4 receptors and astrocytes do not. (A) Cholinergic (ChAT-ir; red) AP neurons express CXCR4 receptors (green); colocalization appears yellow (examples at arrows). (B) Catecholaminergic

    Techniques Used:

    Rhodamine-dextran-covered latex nanospheres injected into the wall of the stomach were used to identify gastric-projecting DMN neurons. Of 264 DMN neurons retrogradely labeled with rhodamine spheres (rhodamine label, red), 239 (i.e. 90%) expressed CXCR4
    Figure Legend Snippet: Rhodamine-dextran-covered latex nanospheres injected into the wall of the stomach were used to identify gastric-projecting DMN neurons. Of 264 DMN neurons retrogradely labeled with rhodamine spheres (rhodamine label, red), 239 (i.e. 90%) expressed CXCR4

    Techniques Used: Injection, Labeling

    ) were used to replicate their observations of CXCR4 receptor expression on neurons and glia widely throughout the cortex and caudate / putamen. These same methods were applied to our investigations
    Figure Legend Snippet: ) were used to replicate their observations of CXCR4 receptor expression on neurons and glia widely throughout the cortex and caudate / putamen. These same methods were applied to our investigations

    Techniques Used: Expressing

    Low-magnification coronal view of caudal medullary hemisections at the level of the calamus scriptorum (levels are 0.0 mm, 0.3 mm anterior and 0.6 mm anterior to calamus) immunohistochemically stained for CXCR4 receptors. CXCR4 receptors are intensely
    Figure Legend Snippet: Low-magnification coronal view of caudal medullary hemisections at the level of the calamus scriptorum (levels are 0.0 mm, 0.3 mm anterior and 0.6 mm anterior to calamus) immunohistochemically stained for CXCR4 receptors. CXCR4 receptors are intensely

    Techniques Used: Staining

    Medial NST showing that virtually all neurons (red, NeuN-ir; left panel) in this region express CXCR4 receptors (green, CXCR4-ir; middle panel), primarily on their proximal dendrites (merged image; right panel). Arrows show some of the clearer examples.
    Figure Legend Snippet: Medial NST showing that virtually all neurons (red, NeuN-ir; left panel) in this region express CXCR4 receptors (green, CXCR4-ir; middle panel), primarily on their proximal dendrites (merged image; right panel). Arrows show some of the clearer examples.

    Techniques Used:

    Phenotypic identification (red, phenotypic-ir; e.g. NOS, TH, GAD-67, OX-42, GFAP and CNPase) of cells in the NST that express CXCR4 receptors (green, CXCR4-ir). Virtually all NOS+, TH+ and GAD-67+ neuronal phenotypes, as well as microglia, also express
    Figure Legend Snippet: Phenotypic identification (red, phenotypic-ir; e.g. NOS, TH, GAD-67, OX-42, GFAP and CNPase) of cells in the NST that express CXCR4 receptors (green, CXCR4-ir). Virtually all NOS+, TH+ and GAD-67+ neuronal phenotypes, as well as microglia, also express

    Techniques Used:

    CXCR4 receptors on vagal motor neurons in the dorsal motor nucleus (DMN) and hypoglossal nucleus are only found on the ChAT phenotype. Note: red, phenotypic identification; green, CXCR4-ir. (A) Low-magnification photomicrograph showing dense CXCR4 receptor
    Figure Legend Snippet: CXCR4 receptors on vagal motor neurons in the dorsal motor nucleus (DMN) and hypoglossal nucleus are only found on the ChAT phenotype. Note: red, phenotypic identification; green, CXCR4-ir. (A) Low-magnification photomicrograph showing dense CXCR4 receptor

    Techniques Used:

    2) Product Images from "A paracrine network regulates the cross-talk between human lung stem cells and the stroma"

    Article Title: A paracrine network regulates the cross-talk between human lung stem cells and the stroma

    Journal: Nature Communications

    doi: 10.1038/ncomms4175

    SDF-1 released by activated LSCs is the key paracrine factor to recruit stromal fibroblasts. ( a ) Protein array showing differential cytokine release by activated LSCs (ALSC) when co-cultured with fibroblasts. ( b ) The mRNA relative levels of different cytokines confirm their upregulation in ALSCs. ( c ) Time-lapse tracing of fibroblast (red) recruitment by LSCs (green) in lung explants. Scale bars, 200 μm. ( d ) Serial sections images captured at day 4 showing expression of SDF-1 (white) and CXCR4 (purple) by LSCs and fibroblasts, respectively. Scale bars, 50 μm. ( e ) Fibroblast migration induced by conditioned medium (CM) from ALSCs (expressing control shRNA) (line 4) is prevented by addition of an anti-SDF-1 antibody (line 5) or by using CM from LSCs lacking SDF-1 (ALSC-KD) (line 6). Addition of SDF-1 recombinant protein into CM from ALSC-KD cells rescued their potential to recruit fibroblasts (line 7). Scale bar, 200 μm. All results ( b and e ) are the mean±s.e.m. of 4–5 triplicate experiments. P
    Figure Legend Snippet: SDF-1 released by activated LSCs is the key paracrine factor to recruit stromal fibroblasts. ( a ) Protein array showing differential cytokine release by activated LSCs (ALSC) when co-cultured with fibroblasts. ( b ) The mRNA relative levels of different cytokines confirm their upregulation in ALSCs. ( c ) Time-lapse tracing of fibroblast (red) recruitment by LSCs (green) in lung explants. Scale bars, 200 μm. ( d ) Serial sections images captured at day 4 showing expression of SDF-1 (white) and CXCR4 (purple) by LSCs and fibroblasts, respectively. Scale bars, 50 μm. ( e ) Fibroblast migration induced by conditioned medium (CM) from ALSCs (expressing control shRNA) (line 4) is prevented by addition of an anti-SDF-1 antibody (line 5) or by using CM from LSCs lacking SDF-1 (ALSC-KD) (line 6). Addition of SDF-1 recombinant protein into CM from ALSC-KD cells rescued their potential to recruit fibroblasts (line 7). Scale bar, 200 μm. All results ( b and e ) are the mean±s.e.m. of 4–5 triplicate experiments. P

    Techniques Used: Protein Array, Cell Culture, Expressing, Migration, shRNA, Recombinant

    3) Product Images from "Chemokines and Glycoprotein120 Produce Pain Hypersensitivity by Directly Exciting Primary Nociceptive Neurons"

    Article Title: Chemokines and Glycoprotein120 Produce Pain Hypersensitivity by Directly Exciting Primary Nociceptive Neurons

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.21-14-05027.2001

    RT-PCR analysis of chemokine receptor expression on neonatal DRG neurons. Results demonstrate the presence of mRNA of CXCR4, CX3CR1, CCR5, and CCR4 in DRG neurons. Lanes 2 and 3 in each panel show PCR products obtained from amplification by primers selected specifically to detect each chemokine receptor ( lane 2 , DRG neurons; lane 3 , rat brain). Lane 1 contains 1 kb plus ladder (Life Technologies); lane 4 indicates no amplification products with H 2 O.
    Figure Legend Snippet: RT-PCR analysis of chemokine receptor expression on neonatal DRG neurons. Results demonstrate the presence of mRNA of CXCR4, CX3CR1, CCR5, and CCR4 in DRG neurons. Lanes 2 and 3 in each panel show PCR products obtained from amplification by primers selected specifically to detect each chemokine receptor ( lane 2 , DRG neurons; lane 3 , rat brain). Lane 1 contains 1 kb plus ladder (Life Technologies); lane 4 indicates no amplification products with H 2 O.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing, Polymerase Chain Reaction, Amplification

    Confocal laser microscopy analysis of CXCR4 expression on HEK 293 cells and DRG neurons. A , Illustrated is immunohistochemical staining for the cloned rat CXCR4 receptor expressed in HEK 293 cells ( a ). Also illustrated are controls in which the primary antibody was not applied ( b ) or HEK 293 cells in which the receptor was not expressed ( c ). B , Staining of cultured rat DRG cells revealed a population of neurons that expressed the CXCR4 receptor ( a ). Note the staining of both the cell soma and terminal varicosities. Staining for the CXCR4 receptor could be blocked by preabsorbing the antiserum with the peptide epitope against which it was raised ( b ). C , Colocalization of substance P and the CXCR4 receptor in rat DRG neurons. a , DRG neuron staining for substance P. b , Two DRG neurons staining for the CXCR4 receptor. c , Overlay of images in a and b showing the colocalization of substance P and the CXCR4 receptor in one neuron ( black arrow ) and the absence of substance P ( white arrow ) in the second CXCR4-stained neuron. D , Colocalization of VR1 and the CXCR4 receptor in rat DRG neurons. a , DRG neuron staining for VR1. b , Staining for CXCR4. c , Overlay of images in a and b showing the colocalization of VR1 and the CXCR4 receptor.
    Figure Legend Snippet: Confocal laser microscopy analysis of CXCR4 expression on HEK 293 cells and DRG neurons. A , Illustrated is immunohistochemical staining for the cloned rat CXCR4 receptor expressed in HEK 293 cells ( a ). Also illustrated are controls in which the primary antibody was not applied ( b ) or HEK 293 cells in which the receptor was not expressed ( c ). B , Staining of cultured rat DRG cells revealed a population of neurons that expressed the CXCR4 receptor ( a ). Note the staining of both the cell soma and terminal varicosities. Staining for the CXCR4 receptor could be blocked by preabsorbing the antiserum with the peptide epitope against which it was raised ( b ). C , Colocalization of substance P and the CXCR4 receptor in rat DRG neurons. a , DRG neuron staining for substance P. b , Two DRG neurons staining for the CXCR4 receptor. c , Overlay of images in a and b showing the colocalization of substance P and the CXCR4 receptor in one neuron ( black arrow ) and the absence of substance P ( white arrow ) in the second CXCR4-stained neuron. D , Colocalization of VR1 and the CXCR4 receptor in rat DRG neurons. a , DRG neuron staining for VR1. b , Staining for CXCR4. c , Overlay of images in a and b showing the colocalization of VR1 and the CXCR4 receptor.

    Techniques Used: Microscopy, Expressing, Immunohistochemistry, Staining, Clone Assay, Cell Culture

    Related Articles

    Incubation:

    Article Title: Mesenchymal Stem Cells Attract Endothelial Progenitor Cells via a Positive Feedback Loop between CXCR2 and CXCR4
    Article Snippet: .. The preparations were then blocked with 5% skimmed milk for 1 hour at room temperature and incubated with the following primary antibodies: anti-CXCR2, anti-CXCR4 (1 : 1000 dilution; Santa Cruz, USA), and anti-pSTAT3 (1 : 2000 dilution; Abcam, USA) overnight at 4°C. ..

    Article Title: miR-330 Regulates Colorectal Cancer Oncogenesis by Targeting BACH1
    Article Snippet: .. Then, they were incubated with primary mouse monoclonal antibodies against BACH1, CXCR4, MMP9, and VEGFR (1:1000, Santa Cruz Biotechnology, Ca) overnight at 4°C. ..

    Staining:

    Article Title: The negative charge of the 343 site is essential for maintaining physiological functions of CXCR4
    Article Snippet: .. Cells were stained with Mouse mAb IgG isotype control (Cell Signaling), anti-CXCR4 (4G10) primary antibody (Santa Cruz Biotechnology) and anti-mouse IgG(H + L) conjugate with Alexa Flour 647 (Cell Signaling) and analyzed by using a four-color FACSCalibur flow cytometer equipped with CELLQUEST PRO software (BD LSRFortessa™ X-20) and analyzed using FlowJo software. ..

    Article Title: Mechanisms of HIV-1 evasion to the antiviral activity of chemokine CXCL12 indicate potential links with pathogenesis
    Article Snippet: .. The following antibodies were used for membrane or intracellular staining: CD45RA-APC (clone T6D11), CCR7-PercP-Vio700 (clone REA546), HLA-DR-APC-Vio770 (clone AC122), CD3-PE-Vio615 (clone REA613), CCR5-PE-Vio770 (clone REA245), CXCR4-PE (clone REA649) (all from Miltenyi Biotec); CD4-BV786 (clone SK3) (from BD Biosciences); CXCR4 (clone 4G10) (Santa Cruz Biotechnology); Unlabeled CXCR4 mAb clones (12G5, 44716, 44717, 44708) (all from R & D Systems); goat anti-mouse secondary antibody-AF647 (from Invitrogen); HIV-1 core antigen-FITC (clone KC57) (from Beckman Coulter). ..

    Flow Cytometry:

    Article Title: The negative charge of the 343 site is essential for maintaining physiological functions of CXCR4
    Article Snippet: .. Cells were stained with Mouse mAb IgG isotype control (Cell Signaling), anti-CXCR4 (4G10) primary antibody (Santa Cruz Biotechnology) and anti-mouse IgG(H + L) conjugate with Alexa Flour 647 (Cell Signaling) and analyzed by using a four-color FACSCalibur flow cytometer equipped with CELLQUEST PRO software (BD LSRFortessa™ X-20) and analyzed using FlowJo software. ..

    Software:

    Article Title: The negative charge of the 343 site is essential for maintaining physiological functions of CXCR4
    Article Snippet: .. Cells were stained with Mouse mAb IgG isotype control (Cell Signaling), anti-CXCR4 (4G10) primary antibody (Santa Cruz Biotechnology) and anti-mouse IgG(H + L) conjugate with Alexa Flour 647 (Cell Signaling) and analyzed by using a four-color FACSCalibur flow cytometer equipped with CELLQUEST PRO software (BD LSRFortessa™ X-20) and analyzed using FlowJo software. ..

    Fluorescence:

    Article Title: Aspirin enhances cisplatin sensitivity of resistant non-small cell lung carcinoma stem-like cells by targeting mTOR-Akt axis to repress migration
    Article Snippet: .. Mean fluorescence intensities of Akt, p-Akt, Cxcr-4, Mdr1, Integrin-α2, Integrin-α5, Integrin-β1, p-Fak (Santa Cruz Biotechnology, Inc.) were determined with respective primary antibodies conjugated with PE as previously described . ..

    Clone Assay:

    Article Title: Mechanisms of HIV-1 evasion to the antiviral activity of chemokine CXCL12 indicate potential links with pathogenesis
    Article Snippet: .. The following antibodies were used for membrane or intracellular staining: CD45RA-APC (clone T6D11), CCR7-PercP-Vio700 (clone REA546), HLA-DR-APC-Vio770 (clone AC122), CD3-PE-Vio615 (clone REA613), CCR5-PE-Vio770 (clone REA245), CXCR4-PE (clone REA649) (all from Miltenyi Biotec); CD4-BV786 (clone SK3) (from BD Biosciences); CXCR4 (clone 4G10) (Santa Cruz Biotechnology); Unlabeled CXCR4 mAb clones (12G5, 44716, 44717, 44708) (all from R & D Systems); goat anti-mouse secondary antibody-AF647 (from Invitrogen); HIV-1 core antigen-FITC (clone KC57) (from Beckman Coulter). ..

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  • 86
    Santa Cruz Biotechnology rabbit anti human cxcr4
    Expression of <t>CXCR4</t> by Western blotting and measurement of reactive oxygen species (ROS) by H 2 DCFDA using fluorescence intensity. ( A ) Different IRB concentrations induced different expression levels of CXCR4. At a concentration of 15 μg/mL, CXCR4 RNA was increased by approximately 2-fold ( n = 8, *: p ≤ 0.05, ***: p ≤ 0.005). ( B ) Different IRB concentrations and IRB incubation time induced changes in ROS levels. After 3 h, 20 μg/mL IRBs clearly generated ROS compared to 0 μg/mL. After 24 h, 15 μg/mL induced a nearly 2-fold increase in ROS levels compared to in the control and 20 μg/mL induced a more than 2-fold increase in ROS levels compared to in the control. H 2 DCFDA and IRBs were added to each well at the same time ( n = 9, *: p ≤ 0.05, ***: p ≤ 0.005).
    Rabbit Anti Human Cxcr4, supplied by Santa Cruz Biotechnology, 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/rabbit anti human cxcr4/product/Santa Cruz Biotechnology
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti human cxcr4 - by Bioz Stars, 2021-09
    86/100 stars
      Buy from Supplier

    86
    Santa Cruz Biotechnology anti cxcr4
    Effect of DAMGO on surface expression of <t>CXCR4</t> in SH-SY5Y cells. Surface expression of CXCR4 in SH-SY5Y cells was assessed by flow cytometry (all experiments repeated twice). ( A ) Untreated = blue; DAMGO 10 μ M = green; CXCL12 (1 h) = red; IgG2B-PE isotype control = grey. ( B ) Cells cotreated with DAMGO+CXCL12 are shown in purple.
    Anti Cxcr4, supplied by Santa Cruz Biotechnology, 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 cxcr4/product/Santa Cruz Biotechnology
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cxcr4 - by Bioz Stars, 2021-09
    86/100 stars
      Buy from Supplier

    95
    Santa Cruz Biotechnology anti cxcr4 antibody
    Interaction between <t>CXCR4</t> and CB2 inhibited CXCR4-mediated functions. Serum-starved PC3 ( A ) or MDA-MB-231 ( B ) cells, both preincubated with calcium dye for 1 h, were treated with SDF1α, AM1241, AMD3100, or SDF1α/AM1241 simultaneously at
    Anti Cxcr4 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cxcr4 antibody/product/Santa Cruz Biotechnology
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti cxcr4 antibody - by Bioz Stars, 2021-09
    95/100 stars
      Buy from Supplier

    Image Search Results


    Expression of CXCR4 by Western blotting and measurement of reactive oxygen species (ROS) by H 2 DCFDA using fluorescence intensity. ( A ) Different IRB concentrations induced different expression levels of CXCR4. At a concentration of 15 μg/mL, CXCR4 RNA was increased by approximately 2-fold ( n = 8, *: p ≤ 0.05, ***: p ≤ 0.005). ( B ) Different IRB concentrations and IRB incubation time induced changes in ROS levels. After 3 h, 20 μg/mL IRBs clearly generated ROS compared to 0 μg/mL. After 24 h, 15 μg/mL induced a nearly 2-fold increase in ROS levels compared to in the control and 20 μg/mL induced a more than 2-fold increase in ROS levels compared to in the control. H 2 DCFDA and IRBs were added to each well at the same time ( n = 9, *: p ≤ 0.05, ***: p ≤ 0.005).

    Journal: International Journal of Molecular Sciences

    Article Title: Enhanced Homing Technique of Mesenchymal Stem Cells Using Iron Oxide Nanoparticles by Magnetic Attraction in Olfactory-Injured Mouse Models

    doi: 10.3390/ijms19051376

    Figure Lengend Snippet: Expression of CXCR4 by Western blotting and measurement of reactive oxygen species (ROS) by H 2 DCFDA using fluorescence intensity. ( A ) Different IRB concentrations induced different expression levels of CXCR4. At a concentration of 15 μg/mL, CXCR4 RNA was increased by approximately 2-fold ( n = 8, *: p ≤ 0.05, ***: p ≤ 0.005). ( B ) Different IRB concentrations and IRB incubation time induced changes in ROS levels. After 3 h, 20 μg/mL IRBs clearly generated ROS compared to 0 μg/mL. After 24 h, 15 μg/mL induced a nearly 2-fold increase in ROS levels compared to in the control and 20 μg/mL induced a more than 2-fold increase in ROS levels compared to in the control. H 2 DCFDA and IRBs were added to each well at the same time ( n = 9, *: p ≤ 0.05, ***: p ≤ 0.005).

    Article Snippet: Membranes were hybridized with rabbit anti-human CXCR4 at 1:1000 (sc-9046; Santa Cruz Biotechnology, Dallas, TX, USA), with SDF-1 (ab18919; Abcam, Cambridge, UK), or anti-β-actin (1:500; Sigma, St. Louis, MO, USA).

    Techniques: Expressing, Western Blot, Fluorescence, Concentration Assay, Incubation, Generated

    Expression of CXCR4 by Western blotting and measurement of reactive oxygen species (ROS) by H 2 DCFDA using fluorescence intensity. ( A ) Different IRB concentrations induced different expression levels of CXCR4. At a concentration of 15 μg/mL, CXCR4 RNA was increased by approximately 2-fold ( n = 8, *: p ≤ 0.05, ***: p ≤ 0.005). ( B ) Different IRB concentrations and IRB incubation time induced changes in ROS levels. After 3 h, 20 μg/mL IRBs clearly generated ROS compared to 0 μg/mL. After 24 h, 15 μg/mL induced a nearly 2-fold increase in ROS levels compared to in the control and 20 μg/mL induced a more than 2-fold increase in ROS levels compared to in the control. H 2 DCFDA and IRBs were added to each well at the same time ( n = 9, *: p ≤ 0.05, ***: p ≤ 0.005).

    Journal: International Journal of Molecular Sciences

    Article Title: Enhanced Homing Technique of Mesenchymal Stem Cells Using Iron Oxide Nanoparticles by Magnetic Attraction in Olfactory-Injured Mouse Models

    doi: 10.3390/ijms19051376

    Figure Lengend Snippet: Expression of CXCR4 by Western blotting and measurement of reactive oxygen species (ROS) by H 2 DCFDA using fluorescence intensity. ( A ) Different IRB concentrations induced different expression levels of CXCR4. At a concentration of 15 μg/mL, CXCR4 RNA was increased by approximately 2-fold ( n = 8, *: p ≤ 0.05, ***: p ≤ 0.005). ( B ) Different IRB concentrations and IRB incubation time induced changes in ROS levels. After 3 h, 20 μg/mL IRBs clearly generated ROS compared to 0 μg/mL. After 24 h, 15 μg/mL induced a nearly 2-fold increase in ROS levels compared to in the control and 20 μg/mL induced a more than 2-fold increase in ROS levels compared to in the control. H 2 DCFDA and IRBs were added to each well at the same time ( n = 9, *: p ≤ 0.05, ***: p ≤ 0.005).

    Article Snippet: Membranes were hybridized with rabbit anti-human CXCR4 at 1:1000 (sc-9046; Santa Cruz Biotechnology, Dallas, TX, USA), with SDF-1 (ab18919; Abcam, Cambridge, UK), or anti-β-actin (1:500; Sigma, St. Louis, MO, USA).

    Techniques: Expressing, Western Blot, Fluorescence, Concentration Assay, Incubation, Generated

    Effect of DAMGO on surface expression of CXCR4 in SH-SY5Y cells. Surface expression of CXCR4 in SH-SY5Y cells was assessed by flow cytometry (all experiments repeated twice). ( A ) Untreated = blue; DAMGO 10 μ M = green; CXCL12 (1 h) = red; IgG2B-PE isotype control = grey. ( B ) Cells cotreated with DAMGO+CXCL12 are shown in purple.

    Journal: Journal of neurovirology

    Article Title: Modulation of neuronal CXCR4 by the μ-opioid agonist DAMGO

    doi: 10.1080/13550280601064798

    Figure Lengend Snippet: Effect of DAMGO on surface expression of CXCR4 in SH-SY5Y cells. Surface expression of CXCR4 in SH-SY5Y cells was assessed by flow cytometry (all experiments repeated twice). ( A ) Untreated = blue; DAMGO 10 μ M = green; CXCL12 (1 h) = red; IgG2B-PE isotype control = grey. ( B ) Cells cotreated with DAMGO+CXCL12 are shown in purple.

    Article Snippet: The following antibodies were used: anti-CXCR4 (H-118 and G-19, raised against amino acids 176 to 293 and N-terminus, respectively) (1:500); anti-MOR (H-80) (1:500) from Santa Cruz Biotech; anti-Akt, anti-phosphoAkt (Ser473), anti-ERK, and anti-phosphoERK from Cell Signaling (1:2,000); anti- β -actin from Sigma-Aldrich (1:5000).

    Techniques: Expressing, Flow Cytometry, Cytometry

    Effect of DAMGO on total and plasma membrane levels of CXCR4 in cortical neurons. CXCR4 protein levels in cortical neurons were studied by Western blot ( top panel ) and confocal microscopy ( A – H ) as described in Methods. As shown by the representative gel in the top panel ( n = 3), cortical neurons were treated with DAMGO up to 48 h. The confocal microscopy studies were preformed on cultures treated with DAMGO (1 μ M) for 24 h. In both sets of experiments, control and DAMGO-treated neurons appeared to have similar levels of CXCR4. For the confocal studies, neurons were stained with an antibody against CXCR4 ( green ) and the nuclear dye Hoechst 33342 ( blue ) as detailed in methods section, and visualized using a 63X 1.4 oil immersion objective mounted on a Leica confocal microscope. Images were collected by using a step size of 0.3 μ m along the z axis (512 × 512; 8 bits/pixel). A and F: Negative control (i.e., without primary antibody). B, D , and G: Control. C, E , and H: DAMGO-treated. A total of 100 cells per treatment were studied from two independent experiments (three coverslips/group).

    Journal: Journal of neurovirology

    Article Title: Modulation of neuronal CXCR4 by the μ-opioid agonist DAMGO

    doi: 10.1080/13550280601064798

    Figure Lengend Snippet: Effect of DAMGO on total and plasma membrane levels of CXCR4 in cortical neurons. CXCR4 protein levels in cortical neurons were studied by Western blot ( top panel ) and confocal microscopy ( A – H ) as described in Methods. As shown by the representative gel in the top panel ( n = 3), cortical neurons were treated with DAMGO up to 48 h. The confocal microscopy studies were preformed on cultures treated with DAMGO (1 μ M) for 24 h. In both sets of experiments, control and DAMGO-treated neurons appeared to have similar levels of CXCR4. For the confocal studies, neurons were stained with an antibody against CXCR4 ( green ) and the nuclear dye Hoechst 33342 ( blue ) as detailed in methods section, and visualized using a 63X 1.4 oil immersion objective mounted on a Leica confocal microscope. Images were collected by using a step size of 0.3 μ m along the z axis (512 × 512; 8 bits/pixel). A and F: Negative control (i.e., without primary antibody). B, D , and G: Control. C, E , and H: DAMGO-treated. A total of 100 cells per treatment were studied from two independent experiments (three coverslips/group).

    Article Snippet: The following antibodies were used: anti-CXCR4 (H-118 and G-19, raised against amino acids 176 to 293 and N-terminus, respectively) (1:500); anti-MOR (H-80) (1:500) from Santa Cruz Biotech; anti-Akt, anti-phosphoAkt (Ser473), anti-ERK, and anti-phosphoERK from Cell Signaling (1:2,000); anti- β -actin from Sigma-Aldrich (1:5000).

    Techniques: Western Blot, Confocal Microscopy, Staining, Microscopy, Negative Control

    Coexpression of MOR and CXCR4 in cortical neurons. Expression of β -tubulin III ( A , green), MOR ( C , red), and CXCR4 ( D , green) in cortical neurons was detected by immunocytochemistry and fluor escence microscopy. Nuclei are stained with Hoechst 33342 ( A-E , blue). Double staining ( E ) shows coexpression of MOR and CXCR4 on individual neurons. Immunoblots ( B ) confirm presence of both receptors in cortical neurons. SH-SY5Y and NIH-3T3 cells were used as positive and negative control, respectively; actin levels were determined to verify protein loading. DAMGO (10 μ M, 5–30 min) increased phosphorylation of ERK1/2 and Akt in neuronal extracts ( F ). Graph shows average data from four experiments ( top: phosphorylated ERK1/2 [P-ERK] or Akt [P-Akt]; bottom: total ERK1/2 or Akt). However, the kinetics of response for both ERK and Akt is not identical among different experiments (which is expected in primary cultures) and this reflected by the data in the graph showing mean ± SEM for ERK ( n = 3) and Akt ( n = 4), respectively. Thus, for statistical analysis purposes, peak responses from these experiments (5′ and 15′) were combined and compared to controls ( P

    Journal: Journal of neurovirology

    Article Title: Modulation of neuronal CXCR4 by the μ-opioid agonist DAMGO

    doi: 10.1080/13550280601064798

    Figure Lengend Snippet: Coexpression of MOR and CXCR4 in cortical neurons. Expression of β -tubulin III ( A , green), MOR ( C , red), and CXCR4 ( D , green) in cortical neurons was detected by immunocytochemistry and fluor escence microscopy. Nuclei are stained with Hoechst 33342 ( A-E , blue). Double staining ( E ) shows coexpression of MOR and CXCR4 on individual neurons. Immunoblots ( B ) confirm presence of both receptors in cortical neurons. SH-SY5Y and NIH-3T3 cells were used as positive and negative control, respectively; actin levels were determined to verify protein loading. DAMGO (10 μ M, 5–30 min) increased phosphorylation of ERK1/2 and Akt in neuronal extracts ( F ). Graph shows average data from four experiments ( top: phosphorylated ERK1/2 [P-ERK] or Akt [P-Akt]; bottom: total ERK1/2 or Akt). However, the kinetics of response for both ERK and Akt is not identical among different experiments (which is expected in primary cultures) and this reflected by the data in the graph showing mean ± SEM for ERK ( n = 3) and Akt ( n = 4), respectively. Thus, for statistical analysis purposes, peak responses from these experiments (5′ and 15′) were combined and compared to controls ( P

    Article Snippet: The following antibodies were used: anti-CXCR4 (H-118 and G-19, raised against amino acids 176 to 293 and N-terminus, respectively) (1:500); anti-MOR (H-80) (1:500) from Santa Cruz Biotech; anti-Akt, anti-phosphoAkt (Ser473), anti-ERK, and anti-phosphoERK from Cell Signaling (1:2,000); anti- β -actin from Sigma-Aldrich (1:5000).

    Techniques: Expressing, Immunocytochemistry, Microscopy, Staining, Double Staining, Western Blot, Negative Control

    Interaction between CXCR4 and CB2 inhibited CXCR4-mediated functions. Serum-starved PC3 ( A ) or MDA-MB-231 ( B ) cells, both preincubated with calcium dye for 1 h, were treated with SDF1α, AM1241, AMD3100, or SDF1α/AM1241 simultaneously at

    Journal: The Journal of Biological Chemistry

    Article Title: Simultaneous Activation of Induced Heterodimerization between CXCR4 Chemokine Receptor and Cannabinoid Receptor 2 (CB2) Reveals a Mechanism for Regulation of Tumor Progression *

    doi: 10.1074/jbc.M115.712661

    Figure Lengend Snippet: Interaction between CXCR4 and CB2 inhibited CXCR4-mediated functions. Serum-starved PC3 ( A ) or MDA-MB-231 ( B ) cells, both preincubated with calcium dye for 1 h, were treated with SDF1α, AM1241, AMD3100, or SDF1α/AM1241 simultaneously at

    Article Snippet: Equal amounts of protein per sample were precleared with rabbit IgG TrueBlot beads (1:50; Santa Cruz Biotechnology) for 2 h at 4 °C and centrifuged at maximal speed, and the cleared lysate was immunoprecipitated with anti-CXCR4 antibody (1:100; Santa Cruz Biotechnology) overnight at 4 °C followed by an incubation with Protein A/G beads for 2 h at 4 °C.

    Techniques: Multiple Displacement Amplification

    Endogenous CXCR4 and CB2 formed heterodimers in cancer cells. Heterodimerization of CXCR4 and CB2 was recognized by incubating cells with primary antibodies raised in two different species and secondary antibodies linked to different DNA oligomers, one

    Journal: The Journal of Biological Chemistry

    Article Title: Simultaneous Activation of Induced Heterodimerization between CXCR4 Chemokine Receptor and Cannabinoid Receptor 2 (CB2) Reveals a Mechanism for Regulation of Tumor Progression *

    doi: 10.1074/jbc.M115.712661

    Figure Lengend Snippet: Endogenous CXCR4 and CB2 formed heterodimers in cancer cells. Heterodimerization of CXCR4 and CB2 was recognized by incubating cells with primary antibodies raised in two different species and secondary antibodies linked to different DNA oligomers, one

    Article Snippet: Equal amounts of protein per sample were precleared with rabbit IgG TrueBlot beads (1:50; Santa Cruz Biotechnology) for 2 h at 4 °C and centrifuged at maximal speed, and the cleared lysate was immunoprecipitated with anti-CXCR4 antibody (1:100; Santa Cruz Biotechnology) overnight at 4 °C followed by an incubation with Protein A/G beads for 2 h at 4 °C.

    Techniques:

    Diminution of endogenous SDF1α did not inhibit endogenous CXCR4/CB2 heterodimerization. MDA-MB-231 ( A ) and PC3 ( E ) cells were transfected with siRNA (Santa Cruz Biotechnology; 100 n m ) targeting SDF1α prior to harvesting for Duolink. Transfected

    Journal: The Journal of Biological Chemistry

    Article Title: Simultaneous Activation of Induced Heterodimerization between CXCR4 Chemokine Receptor and Cannabinoid Receptor 2 (CB2) Reveals a Mechanism for Regulation of Tumor Progression *

    doi: 10.1074/jbc.M115.712661

    Figure Lengend Snippet: Diminution of endogenous SDF1α did not inhibit endogenous CXCR4/CB2 heterodimerization. MDA-MB-231 ( A ) and PC3 ( E ) cells were transfected with siRNA (Santa Cruz Biotechnology; 100 n m ) targeting SDF1α prior to harvesting for Duolink. Transfected

    Article Snippet: Equal amounts of protein per sample were precleared with rabbit IgG TrueBlot beads (1:50; Santa Cruz Biotechnology) for 2 h at 4 °C and centrifuged at maximal speed, and the cleared lysate was immunoprecipitated with anti-CXCR4 antibody (1:100; Santa Cruz Biotechnology) overnight at 4 °C followed by an incubation with Protein A/G beads for 2 h at 4 °C.

    Techniques: Multiple Displacement Amplification, Transfection

    Simultaneous SDF1α/AM1241 treatment induced physical association between CXCR4 and CB2. A , PC3 cells were treated for 15 min with SDF1α, AM1241, or SDF1α/AM1241 simultaneously. Cells were lysed and then incubated with anti-CXCR4

    Journal: The Journal of Biological Chemistry

    Article Title: Simultaneous Activation of Induced Heterodimerization between CXCR4 Chemokine Receptor and Cannabinoid Receptor 2 (CB2) Reveals a Mechanism for Regulation of Tumor Progression *

    doi: 10.1074/jbc.M115.712661

    Figure Lengend Snippet: Simultaneous SDF1α/AM1241 treatment induced physical association between CXCR4 and CB2. A , PC3 cells were treated for 15 min with SDF1α, AM1241, or SDF1α/AM1241 simultaneously. Cells were lysed and then incubated with anti-CXCR4

    Article Snippet: Equal amounts of protein per sample were precleared with rabbit IgG TrueBlot beads (1:50; Santa Cruz Biotechnology) for 2 h at 4 °C and centrifuged at maximal speed, and the cleared lysate was immunoprecipitated with anti-CXCR4 antibody (1:100; Santa Cruz Biotechnology) overnight at 4 °C followed by an incubation with Protein A/G beads for 2 h at 4 °C.

    Techniques: Incubation

    SDF1α and AM1241 did not compete for binding to CXCR4. MDA-MB-231 ( A and B ) or 293T cells ( A ) were plated in 96-well puncher plates and grown to a density of 75% prior to the experiment. To assign absolute affinity of each ligand for CXCR4 receptor,

    Journal: The Journal of Biological Chemistry

    Article Title: Simultaneous Activation of Induced Heterodimerization between CXCR4 Chemokine Receptor and Cannabinoid Receptor 2 (CB2) Reveals a Mechanism for Regulation of Tumor Progression *

    doi: 10.1074/jbc.M115.712661

    Figure Lengend Snippet: SDF1α and AM1241 did not compete for binding to CXCR4. MDA-MB-231 ( A and B ) or 293T cells ( A ) were plated in 96-well puncher plates and grown to a density of 75% prior to the experiment. To assign absolute affinity of each ligand for CXCR4 receptor,

    Article Snippet: Equal amounts of protein per sample were precleared with rabbit IgG TrueBlot beads (1:50; Santa Cruz Biotechnology) for 2 h at 4 °C and centrifuged at maximal speed, and the cleared lysate was immunoprecipitated with anti-CXCR4 antibody (1:100; Santa Cruz Biotechnology) overnight at 4 °C followed by an incubation with Protein A/G beads for 2 h at 4 °C.

    Techniques: Binding Assay, Multiple Displacement Amplification

    Simultaneous SDF1α/AM1241 treatment inhibited internalization of CXCR4. One million MDA-MB-231 cells were serum-starved for 24 h prior to treatment with SDF1α, AM1241, or SDF1α/AM1241 simultaneously for 10 min. Cells were detached

    Journal: The Journal of Biological Chemistry

    Article Title: Simultaneous Activation of Induced Heterodimerization between CXCR4 Chemokine Receptor and Cannabinoid Receptor 2 (CB2) Reveals a Mechanism for Regulation of Tumor Progression *

    doi: 10.1074/jbc.M115.712661

    Figure Lengend Snippet: Simultaneous SDF1α/AM1241 treatment inhibited internalization of CXCR4. One million MDA-MB-231 cells were serum-starved for 24 h prior to treatment with SDF1α, AM1241, or SDF1α/AM1241 simultaneously for 10 min. Cells were detached

    Article Snippet: Equal amounts of protein per sample were precleared with rabbit IgG TrueBlot beads (1:50; Santa Cruz Biotechnology) for 2 h at 4 °C and centrifuged at maximal speed, and the cleared lysate was immunoprecipitated with anti-CXCR4 antibody (1:100; Santa Cruz Biotechnology) overnight at 4 °C followed by an incubation with Protein A/G beads for 2 h at 4 °C.

    Techniques: Multiple Displacement Amplification

    CXCR4/CB2 heterodimer modulated ERK1/2 phosphorylation. A , serum-starved MDA-MB-231 cells were treated with SDF1α, AM1241, or SDF1α/AM1241 simultaneously for 15 min. Whole protein lysates were isolated prior to separation by 10% SDS-PAGE

    Journal: The Journal of Biological Chemistry

    Article Title: Simultaneous Activation of Induced Heterodimerization between CXCR4 Chemokine Receptor and Cannabinoid Receptor 2 (CB2) Reveals a Mechanism for Regulation of Tumor Progression *

    doi: 10.1074/jbc.M115.712661

    Figure Lengend Snippet: CXCR4/CB2 heterodimer modulated ERK1/2 phosphorylation. A , serum-starved MDA-MB-231 cells were treated with SDF1α, AM1241, or SDF1α/AM1241 simultaneously for 15 min. Whole protein lysates were isolated prior to separation by 10% SDS-PAGE

    Article Snippet: Equal amounts of protein per sample were precleared with rabbit IgG TrueBlot beads (1:50; Santa Cruz Biotechnology) for 2 h at 4 °C and centrifuged at maximal speed, and the cleared lysate was immunoprecipitated with anti-CXCR4 antibody (1:100; Santa Cruz Biotechnology) overnight at 4 °C followed by an incubation with Protein A/G beads for 2 h at 4 °C.

    Techniques: Multiple Displacement Amplification, Isolation, SDS Page