Invitrogen

CXCR4 Polyclonal Antibody

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

CXCR4 Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of CXCR4 (green) showing staining in the cytoplasm of Hela 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 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.

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1:1000 - 1:5000

View 1 publication 1 publication

Immunohistochemistry (IHC)

View 3 publications 3 publications

Immunohistochemistry (Paraffin) (IHC (P))

1:1000 - 1:5000

Immunocytochemistry (ICC)

1:1000 - 1:5000

Immunofluorescence (IF)

1:1000 - 1:5000

Flow Cytometry (Flow)

1:1000 - 1:5000

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.

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

Molecular Function: cell adhesion molecule cytokine receptor defense/immunity protein hydrolase immunoglobulin receptor superfamily immunoglobulin superfamily cell adhesion molecule phosphatase protein phosphatase receptor