Invitrogen

CXCR4 Polyclonal Antibody

Product Image Gallery

CXCR4 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of CXCR4 (green) showing staining in the cytoplasm of U251 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CXCR4 polyclonal antibody (Product # PA3-305) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4°C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. Actin was stained using Alexa Fluor 554 (red) and nuclei were stained with Hoechst or DAPI (blue). Images were taken at a magnification of 60x.

CXCR4 Antibody in Immunohistochemistry (IHC) — Immunostaining of CXCR4 in human cervical carcinoma tissue sections.

CXCR4 Antibody in Western Blot (WB) — Western blot analysis of CXCR4 was performed by loading 25 µg of Ramos, Raji, and Jurkat whole cell lysates or membrane protein fractions, generated using IP Lysis Buffer (Product # 87787) and Mem-PER Plus Membrane Protein Extraction Kit (Product # 89842), respectively, and 10 µL of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a Novex® 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a PVDF membrane using the G2 Blotter (Product # 62288), and blocked with 5% milk in TBST for at least 1 hour at room temperature. CXCR4 was detected at ~60 kD and 72 kD using CXCR4 polyclonal antiserum at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000 for at least 30 minutes. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076). NOTE: The presence of two distinct bands with this antibody likely represents the detection of two different isoforms and/or post-translationally modified forms of CXCR4, many of which have been described in the literature. For optimal detection of CXCR4 by Western blot, it is recommended to first use a membrane extraction buffer for efficient protein isolation.

CXCR4 Antibody in Flow Cytometry (Flow) — Flow cytometry analysis of CXCR4 on Ramos cells. Cells were fixed, permeabilized, and then blocked with a 1:100 dilution of normal goat serum for at least 30 minutes on ice. Cells were stained with CXCR4 polyclonal antiserum (Product # PA3-305, red histogram) or with normal rabbit serum (blue histogram) at a dilution of 1:500. After incubation of the primary antibody for at least 1 hour on ice, the cells were stained with a DyLight 488-conjugated goat anti-rabbit IgG secondary antibody (Product # 35552) for at least 30 minutes on ice. A representative 10,000 cells were acquired for each sample.

CXCR4 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Figure 1 A model of 5-FU submyeloablation to characterize the bone marrow cell trafficking into endometriotic lesions in mice. Schematic model: Treatment for bone marrow conditioning (BMC) started on day -36 before uterine fragment grafting and 6 days before bone marrow transplantation (BMT) (A). Ectopic lesions were harvested 15 and 30 days (each group n = 6) after grafting uterine fragments into the peritoneal. Representative images of flow cytometry analysis of ectopic lesion cells on day 15 and 30 after uterine fragment grafting demonstrating the percentage of GFP-positive cells in endometriosis. B, Control group (n = 6 mice), represented by GFP-negative endometriosis lesions, was used to set a gate. Fluorescence confocal microscopy analysis of mouse endometriosis sections (C, D and E). Representative images of IF and quantitative analyses of cells expressing CXCR4, CXCR7 or CXCL12 and GFP, as a per cent of total endometriosis cells. Murine lesions were stained with anti-GFP antibody (green) and co-stained with CXCR4 antibody (red) (C), with CXCR7 antibody (red) (D) and with CXCL12 antibody (red) (E). Nuclei were stained by DAPI and are shown in blue. BMC, Bone morrow conditioning; BMT, Bone morrow transplantation; PLC, percentage of labelled cells. Results shown as mean +- SE. * P < .05 vs vehicle. Scale bar: 20 mum

Product Details

PA3-305

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000-1:5,000	View 1 publication 1 publication
Immunohistochemistry (IHC)	-	View 4 publications 4 publications
Immunohistochemistry (Paraffin) (IHC (P))	1:1,000-1:5,000	View 2 publications 2 publications
Immunocytochemistry (ICC/IF)	1:1,000-1:5,000	View 3 publications 3 publications
Flow Cytometry (Flow)	1:1,000-1:5,000	-

Product Specifications
Species Reactivity	Human, Mouse
Published species	Human, Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Synthetic peptide corresponding to residues K(338) G K R G G H S S V S T E S E S S S F H S S (359) of human CXCR4 receptor protein.
Conjugate	Unconjugated
Form	Liquid
Concentration	Conc. Not Determined
Storage buffer	whole serum
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2091817

Product Specific Information

Several different molecular weight isoforms of CXCR4 have been described in the literature, which vary depending on cell type tested and extraction buffer used. Using Ramos, Raji, and Jurkat membrane fractions, two different MW forms of CXCR4 were detected with PA3-305 by Western blot, between 55-72kD.

It is recommended to use a membrane isolation buffer/kit for extracting CXCR4 for Western blot analysis. Cellular membrane fractions were isolated for Western blot using Mem-PER Plus (Product # 89842).

Target Information

CXCR4 (Fusin, LESTR, HUMSTR) is a member of the G-protein-coupled chemokine receptor family with seven membrane-spanning domains, and functions as a co-receptor for X4 HIV-1 entry into CD4+ cells. CXCR4 has been proved to be the co-receptor for HIV-2's binding to CD4 through envelope glycoprotein gp 120 and promotes Env-mediated fusion of the virus. In other cases, CXCR4 can even function as the only receptor for HIV-2's binding to the CD4 host cells. CXCR4 is the receptor for the C-X-C chemokine CXCL12/SDF-1 that transduces a signal by increasing intracellular calcium ion levels and enhancing MAPK1/MAPK3 activation. CXCR4 acts as a receptor for extracellular ubiquitin; leading to enhanced intracellular calcium ions and reduced cellular cAMP levels. Also, CXCR4 is involved in hematopoiesis and in cardiac ventricular septum formation, and plays an essential role in vascularization of the gastrointestinal tract, probably by regulating vascular branching and/or remodeling processes in endothelial cells. In the CNS, CXCR4 could mediate hippocampal-neuron survival and is involved in cerebellar development. CXCR4 binds bacterial lipopolysaccharide (LPS) and mediates LPS-induced inflammatory response, including TNF secretion by monocytes. Antibodies to CXCR4 block HIV-1 and HIV-2 fusion and infection of human target cells. The amino-terminal domain and the second extracellular loop of CXCR4 serve as HIV biding sites.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

CXCL12 enhances pregnancy outcome via improvement of endometrial receptivity in mice.

Scientific reports

Koo HS,Yoon MJ,Hong SH,Ahn J,Cha H,Lee D,Ko JE,Kwon H,Choi DH,Lee KA,Ko JJ,Kang YJ

Published figure using CXCR4 polyclonal antibody (Product # PA3-305) in Immunocytochemistry

Thu Apr 01 00:00:00 EDT 2021

Enhanced anti-inflammatory effects of mesenchymal stromal cells mediated by the transient ectopic expression of CXCR4 and IL10.

Stem cell research & therapy

Hervás-Salcedo R,Fernández-García M,Hernando-Rodríguez M,Quintana-Bustamante O,Segovia JC,Alvarez-Silva M,García-Arranz M,Minguez P,Del Pozo V,de Alba MR,García-Olmo D,Ayuso C,Lamana ML,Bueren JA,Yañez RM

Published figure using CXCR4 polyclonal antibody (Product # PA3-305) in Immunocytochemistry

Fri Feb 12 00:00:00 EST 2021

CXCL12-CXCR4 Interplay Facilitates Palatal Osteogenesis in Mice.

Frontiers in cell and developmental biology

Verheijen N,Suttorp CM,van Rheden REM,Regan RF,Helmich MPAC,Kuijpers-Jagtman AM,Wagener FADTG

PA3-305 was used in Immunohistochemistry to provide novel evidence that CXCL12-CXCR4 interplay facilitates palatal osteogenesis and palatal fusion in mice.

Mon Sep 28 00:00:00 EDT 2020

CXCL12-CXCR4 Interplay Facilitates Palatal Osteogenesis in Mice.

Frontiers in cell and developmental biology

Verheijen N,Suttorp CM,van Rheden REM,Regan RF,Helmich MPAC,Kuijpers-Jagtman AM,Wagener FADTG

PA3-305 was used in Immunohistochemistry to provide novel evidence that CXCL12-CXCR4 interplay facilitates palatal osteogenesis and palatal fusion in mice.

Mon Sep 28 00:00:00 EDT 2020

Oral lichen planus interactome reveals CXCR4 and CXCL12 as candidate therapeutic targets.

Scientific reports

Rivera C,Crisóstomo MF,Peña C,González-Díaz P,González-Arriagada WA

PA3-305 was used in Immunohistochemistry to support the pathological role of chemokines and chemokine receptors in oral lichen planus.

Wed Mar 25 00:00:00 EDT 2020

Oral lichen planus interactome reveals CXCR4 and CXCL12 as candidate therapeutic targets.

Scientific reports

Rivera C,Crisóstomo MF,Peña C,González-Díaz P,González-Arriagada WA

PA3-305 was used in Immunohistochemistry to support the pathological role of chemokines and chemokine receptors in oral lichen planus.

Wed Mar 25 00:00:00 EDT 2020

CXCR4 or CXCR7 antagonists treat endometriosis by reducing bone marrow cell trafficking.

Journal of cellular and molecular medicine

Pluchino N,Mamillapalli R,Shaikh S,Habata S,Tal A,Gaye M,Taylor HS

PA3-305 was used in Immunohistochemistry (Paraffin) to propose that CXCR4 and CXCR7 antagonists can be used as potential novel, non-hormonal therapies for endometriosis.

Sat Feb 01 00:00:00 EST 2020

CXCR4 or CXCR7 antagonists treat endometriosis by reducing bone marrow cell trafficking.

Journal of cellular and molecular medicine

Pluchino N,Mamillapalli R,Shaikh S,Habata S,Tal A,Gaye M,Taylor HS

PA3-305 was used in Immunohistochemistry (Paraffin) to propose that CXCR4 and CXCR7 antagonists can be used as potential novel, non-hormonal therapies for endometriosis.

Sat Feb 01 00:00:00 EST 2020

CXCR4 or CXCR7 antagonists treat endometriosis by reducing bone marrow cell trafficking.

Journal of cellular and molecular medicine

Pluchino N,Mamillapalli R,Shaikh S,Habata S,Tal A,Gaye M,Taylor HS

PA3-305 was used in Immunohistochemistry (Paraffin) to propose that CXCR4 and CXCR7 antagonists can be used as potential novel, non-hormonal therapies for endometriosis.

Sat Feb 01 00:00:00 EST 2020

Prognostic value of immunoexpression of CCR4, CCR5, CCR7 and CXCR4 in squamous cell carcinoma of tongue and floor of the mouth.

Medicina oral, patologia oral y cirugia bucal

Domingueti CB,Janini JB,Paranaíba LM,Lozano-Burgos C,Olivero P,González-Arriagada WA

PA3-305 was used in Immunohistochemistry to evaluate the prognostic value of the immunoexpression of chemokines in squamous cell carcinoma of tongue and floor of the mouth.

Wed May 01 00:00:00 EDT 2019

Prognostic value of immunoexpression of CCR4, CCR5, CCR7 and CXCR4 in squamous cell carcinoma of tongue and floor of the mouth.

Medicina oral, patologia oral y cirugia bucal

Domingueti CB,Janini JB,Paranaíba LM,Lozano-Burgos C,Olivero P,González-Arriagada WA

PA3-305 was used in Immunohistochemistry to evaluate the prognostic value of the immunoexpression of chemokines in squamous cell carcinoma of tongue and floor of the mouth.

Wed May 01 00:00:00 EDT 2019

CXCR4 receptor blockage reduces the contribution of tumor and stromal cells to the metastatic growth in the liver.

Oncology reports

Benedicto A,Romayor I,Arteta B

PA3-305 was used in Western Blotting to investigate the effectiveness of CXCR4 receptor blockage, and its mechanisms, in reducing metastatic growth in the liver.

Sun Apr 01 00:00:00 EDT 2018

Locoregional Confinement and Major Clinical Benefit of ¹⁸⁸Re-Loaded CXCR4-Targeted Nanocarriers in an Orthotopic Human to Mouse Model of Glioblastoma.

Theranostics

Séhédic D,Chourpa I,Tétaud C,Griveau A,Loussouarn C,Avril S,Legendre C,Lepareur N,Wion D,Hindré F,Davodeau F,Garcion E

PA3-305 was used in Immunohistochemistry to investigate the efficacy of a novel system of local brain internal radiation therapy, combining a radiopharmaceutical with an immuno-nanoparticle.

Mon Mar 19 00:00:00 EDT 2018

CXCR4 regulates interneuron migration in the developing neocortex.

The Journal of neuroscience : the official journal of the Society for Neuroscience

Stumm RK,Zhou C,Ara T,Lazarini F,Dubois-Dalcq M,Nagasawa T,Höllt V,Schulz S

PA3-305 was used in immunohistochemistry to investigate the important role of CXCR4 in interneuron migration

Sun Jun 15 00:00:00 EDT 2003

Bioinformatics

Protein Aliases: C Cmotif chemokine; C X C motif chemokine; C-X-C chemokine receptor type 4; CC motif chemokine; CCmotif chemokine; CD antigen CD184; CD184; CD184 antigen; chemokine (C-X-C motif) receptor 4; chemokine (C-X-C) receptor 4; chemokine receptor; chemokine receptor 4; CXC; CXC motif chemokine; CXC-R4; CXCR-4; FB22; Fusin; HM89; HUMSTR; LAP-3; LCR1; LESTR; Leukocyte-derived seven transmembrane domain receptor; leukocyte-expressed seven-transmembrane-domain; Lipopolysaccharide-associated protein 3; LPS-associated protein 3; neuropeptide Y receptor Y3; neuropeptide Y3 receptor; NPYRL; PB-CKR; Pre-B-cell-derived chemokine receptor; SDF-1 receptor; seven transmembrane helix receptor; seven-transmembrane-segment receptor, spleen; Stromal cell-derived factor 1 receptor

Gene Aliases: b2b220Clo; CD184; Cmkar4; CXCR4; D2S201E; FB22; HM89; HSY3RR; LAP-3; LAP3; LCR1; LESTR; NPY3R; NPYR; NPYRL; NPYY3R; PB-CKR; PBSF/SDF-1; Sdf1r; WHIM; WHIMS

UniProt ID: (Human) P61073, (Mouse) P70658

Entrez Gene ID: (Human) 7852, (Mouse) 12767

Function(s)