Key features and details
- Rabbit polyclonal to CXCR4 (phospho S339)
- Suitable for: WB, IHC-P, ICC/IF
- Reacts with: Mouse, Human
- Isotype: IgG
Product nameAnti-CXCR4 (phospho S339) antibody
See all CXCR4 primary antibodies
DescriptionRabbit polyclonal to CXCR4 (phospho S339)
Specificityab74012 detects endogenous levels of CXCR4 only when phosphorylated at serine 339 (Mouse: Ser 346).
Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Mouse, Human
Synthetic phosphopeptide derived from human CXCR4 around the phosphorylation site of serine 339 (H-S-SP-V-S).
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 50% Glycerol, 0.87% Sodium chloride, PBS
Without Mg2+ and Ca2+
Concentration information loading...
PurityImmunogen affinity purified
Purification notesab74012 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
Our Abpromise guarantee covers the use of ab74012 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500 - 1/1000. Detects a band of approximately 45 kDa (predicted molecular weight: 40 kDa).|
|IHC-P||1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||1/500 - 1/1000.|
FunctionReceptor for the C-X-C chemokine CXCL12/SDF-1 that transduces a signal by increasing intracellular calcium ions levels and enhancing MAPK1/MAPK3 activation. Acts as a receptor for extracellular ubiquitin; leading to enhance intracellular calcium ions and reduce cellular cAMP levels. Involved in haematopoiesis and in cardiac ventricular septum formation. Plays also an essential role in vascularization of the gastrointestinal tract, probably by regulating vascular branching and/or remodeling processes in endothelial cells. Could be involved in cerebellar development. In the CNS, could mediate hippocampal-neuron survival. Acts as a coreceptor (CD4 being the primary receptor) for HIV-1 X4 isolates and as a primary receptor for some HIV-2 isolates. Promotes Env-mediated fusion of the virus.
Tissue specificityExpressed in numerous tissues, such as peripheral blood leukocytes, spleen, thymus, spinal cord, heart, placenta, lung, liver, skeletal muscle, kidney, pancreas, cerebellum, cerebral cortex and medulla (in microglia as well as in astrocytes), brain microvascular, coronary artery and umbilical cord endothelial cells. Isoform 1 is predominant in all tissues tested.
Involvement in diseaseDefects in CXCR4 are a cause of WHIM syndrome (WHIM) [MIM:193670]; also known as warts, hypogammaglobulinemia, infections and myelokathexis. WHIM syndrome is an immunodeficiency disease characterized by neutropenia, hypogammaglobulinemia and extensive human papillomavirus (HPV) infection. Despite the peripheral neutropenia, bone marrow aspirates from affected individuals contain abundant mature myeloid cells, a condition termed myelokathexis.
Sequence similaritiesBelongs to the G-protein coupled receptor 1 family.
DomainThe amino-terminus is critical for ligand binding. Residues in all four extracellular regions contribute to HIV-1 coreceptor activity.
modificationsPhosphorylated on agonist stimulation. Rapidly phosphorylated on serine and threonine residues in the C-terminal. Phosphorylation at Ser-324 and Ser-325 leads to recruitment of ITCH, ubiquitination and protein degradation.
Ubiquitinated by ITCH at the cell membrane on agonist stimulation. The ubiquitin-dependent mechanism, endosomal sorting complex required for transport (ESCRT), then targets CXCR4 for lysosomal degradation. This process is dependent also on prior Ser-/Thr-phosphorylation in the C-terminal of CXCR4. Also binding of ARRB1 to STAM negatively regulates CXCR4 sorting to lysosomes though modulating ubiquitination of SFR5S.
Sulfation on Tyr-21 is required for efficient binding of CXCL12/SDF-1alpha and promotes its dimerization.
O- and N-glycosylated. Asn-11 is the principal site of N-glycosylation. There appears to be very little or no glycosylation on Asn-176. N-glycosylation masks coreceptor function in both X4 and R5 laboratory-adapted and primary HIV-1 strains through inhibiting interaction with their Env glycoproteins. The O-glycosylation chondroitin sulfate attachment does not affect interaction with CXCL12/SDF-1alpha nor its coreceptor activity.
Cellular localizationCell membrane. In unstimulated cells, diffuse pattern on plasma membrane. On agonist stimulation, colocalizes with ITCH at the plasma membrane where it becomes ubiquitinated.
- Information by UniProt
- C-X-C chemokine receptor type 4 antibody
- CD184 antibody
- CD184 antigen antibody
All lanes : Anti-CXCR4 (phospho S339) antibody (ab74012) at 1/500 dilution
Lane 1 : Extracts from HUVEC cells
treated with etoposide (25uM, 24hours)
Lane 2 : Extracts from HUVEC cells
treated with etoposide (25uM, 24hours) with the immunising phosphopeptide at 5 µg
Lysates/proteins at 5 µg per lane.
Predicted band size: 40 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Immunohistochemistry analysis of paraffin-embedded human brain carcinoma tissue using ab74012, at 1/50 dilution, in the presence (right panel) or absence (left panel) of the immunising phosphopeptide.
Immunofluorescence analysis of HeLa cells using ab74012, at 1/500 dilution, in the presence (right panel) or absence (left panel) of the immunising phosphopeptide.
ab74012 has been referenced in 17 publications.
- Liu J et al. Long Non-Coding RNA FEZF1-AS1 Modulates CXCR4 to Promote Cell Proliferation, Warburg Effect and Suppress Cell Apoptosis in Osteosarcoma by Sponging miR-144. Onco Targets Ther 13:2899-2910 (2020). PubMed: 32308422
- Chen Y et al. CUDC-907 blocks multiple pro-survival signals and abrogates microenvironment protection in CLL. J Cell Mol Med 23:340-348 (2019). PubMed: 30353642
- Cunha F et al. Electrical Stimulation Directs Migration, Enhances and Orients Cell Division and Upregulates the Chemokine Receptors CXCR4 and CXCR2 in Endothelial Cells. J Vasc Res 56:39-53 (2019). PubMed: 30995642
- Cornelison RC et al. Convective forces increase CXCR4-dependent glioblastoma cell invasion in GL261 murine model. Sci Rep 8:17057 (2018). PubMed: 30451884
- Lefort S et al. CXCR4 inhibitors could benefit to HER2 but not to triple-negative breast cancer patients. Oncogene 36:1211-1222 (2017). PubMed: 27669438