Western blot analysis of CXCR4 was performed by loading 25ug of Ramos, Raji, and Jurkat whole cell lysates, and 10ul 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 ~60kD using a CXCR4 monoclonal antibody at a concentration of 2ug/ml in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) 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 a 60kD band likely represents an isoform or post-translationally modified form of CXCR4, many of which have been described in the literature.
|Tested species reactivity||Human|
|Published species reactivity||Hamster, Human, Not Applicable|
|Host / Isotype||Mouse / IgG2a, kappa|
|Immunogen||SIV-infected SupT1 cells injected into Balb/c mice.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/ml|
|Immunofluorescence (IF)||Assay Dependent|
|Western Blot (WB)||2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Miscellaneous PubMed (MISC)||See 3 publications below|
|Immunohistochemistry (IHC)||See 1 publications below|
|Immunohistochemistry (Paraffin) (IHC (P))||See 4 publications below|
|Immunohistochemistry (Paraffin, Frozen) (IHC (P, F))||See 1 publications below|
|Flow Cytometry (Flow)||See 1 publications below|
|Immunofluorescence (IF)||See 1 publications below|
By Western blot, 35-8800 detects a band at ~60kD, which likely represents an isoform or post-translationally modified form of CXCR4, many of which have been described in the literature.
Human immunodeficiency virus (HIV) and related viruses require coreceptors, in addition to CD4, to infect target cells. Some G protein-coupled receptors including CCR5, CXCR4, CCR3, CCR2b and CCR8 in the chemokine receptor family, and four new human molecules GPR15, STRL33, GPR1 and V28 were recently identified as HIV coreceptors. Among them, CXCR4 (fusin, LESTR or HUMSTR) is a principal coreceptor for T-cell tropic strains of HIV-1 fusion and entry of human white blood cells. CXCR4 is also required for the infection by dual-tropic strains of HIV-1 and mediates CD-4 independent infection by HIV-2. The a-chemokine SDF-1 is the ligand for CXCR4 and prevents infection by T-tropic HIV-1. CXCR4 associates with the surface CD4-gp120 complex before HIV enters target cells. CXCR4 messenger RNA levels correlated with HIV-1 permissiveness in diverse human cell types. 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.
Pantethine Alters Lipid Composition and Cholesterol Content of Membrane Rafts, With Down-Regulation of CXCL12-Induced T Cell Migration.
35-8800 was used in immunocytochemistry to examine the effects of pantethine on membrane lipid rafts.
|van Gijsel-Bonnello M,Acar N,Molino Y,Bretillon L,Khrestchatisky M,de Reggi M,Gharib B||Journal of cellular physiology (230:2415)||2015|
Immunohistochemical prediction of brain metastases in patients with advanced breast cancer: the role of Rad51.
35-8800 was used in immunohistochemistry to correlate expression of proteins in the primary tumor with the risk of brain metastases in patients with metastatic breast cancer.
|Sosi¿ska-Mielcarek K,Duchnowska R,Winczura P,Badzio A,Majewska H,Lakomy J,P¿ksa R,Pieczy¿ska B,Radecka B,D¿bska S,Biernat W,Jassem J||Breast (Edinburgh, Scotland) (22:1178)||2013|
Co-culture of hematopoietic stem cells with mesenchymal stem cells increases VCAM-1-dependent migration of primitive hematopoietic stem cells.
35-8800 was used in flow cytometry to investigate how mesenchymal stem cells improve hematopoietic stem cell transplantability.
|Perdomo-Arciniegas AM,Vernot JP||International journal of hematology (94:525)||2011|
Immunohistochemical Predictors of Bone Metastases in Breast Cancer Patients.
35-8800 was used in immunohistochemistry to evaluate the potential protein markers of bone metastases in breast cancer patients.
|Winczura P,Sosi¿ska-Mielcarek K,Duchnowska R,Badzio A,Lakomy J,Majewska H,P¿ksa R,Pieczy¿ska B,Radecka B,D¿bska-Szmich S,Adamowicz K,Biernat W,Jassem J||Pathology oncology research : POR (21:1229)||2015|
Immunohistochemical analysis of cancer stem cell markers in pancreatic adenocarcinoma patients after neoadjuvant chemoradiotherapy.
35-8800 was used in immunohistochemistry - paraffin section to investigate the effect of neoadjuvant chemoradiotherapy on pancreatic adenocarcinoma cancer stem cell markers
|Mizukami T,Kamachi H,Mitsuhashi T,Tsuruga Y,Hatanaka Y,Kamiyama T,Matsuno Y,Taketomi A||BMC cancer (14:null)||2014|
Brain metastasis in renal cancer patients: metastatic pattern, tumour-associated macrophages and chemokine/chemoreceptor expression.
35-8800 was used in immunohistochemistry - paraffin section to discuss the role of chemokines in brain metastasis observed in patients with renal cell cancer
|Wyler L,Napoli CU,Ingold B,Sulser T,Heikenwälder M,Schraml P,Moch H||British journal of cancer (110:686)||2014|
Comparative analysis of tumorbiology and CD133 positivity in primary and recurrent pancreatic ductal adenocarcinoma.
35-8800 was used in immunohistochemistry - paraffin section to measure the expression of CXCR4, actinin-4, and CD133 in primary and recurrent pancreatic ductal adenocarcinoma
|Welsch T,Keleg S,Bergmann F,Degrate L,Bauer S,Schmidt J||Clinical and experimental metastasis (26:701)||2009|
Gene expression profiling identifies genes associated with invasive intraductal papillary mucinous neoplasms of the pancreas.
35-8800 was used in immunohistochemistry - paraffin section to examine the gene expression profiles of intraductal papillary mucinous neoplasms
|Sato N,Fukushima N,Maitra A,Iacobuzio-Donahue CA,van Heek NT,Cameron JL,Yeo CJ,Hruban RH,Goggins M||The American journal of pathology (164:903)||2004|
|Human||Not Cited||Increased levels of active c-Src distinguish invasive from in situ lobular lesions.||Zou D,Yoon HS,Anjomshoaa A,Perez D,Fukuzawa R,Guilford P,Humar B||Breast cancer research : BCR (11:null)||2009|
||CD4-independent infection by HIV-2 is mediated by fusin/CXCR4.||Endres MJ,Clapham PR,Marsh M,Ahuja M,Turner JD,McKnight A,Thomas JF,Stoebenau-Haggarty B,Choe S,Vance PJ,Wells TN,Power CA,Sutterwala SS,Doms RW,Landau NR,Hoxie JA||Cell (87:745)||1996|
|Hamster||Not Cited||CD4-independent infection by HIV-2 is mediated by fusin/CXCR4.||Endres MJ,Clapham PR,Marsh M,Ahuja M,Turner JD,McKnight A,Thomas JF,Stoebenau-Haggarty B,Choe S,Vance PJ,Wells TN,Power CA,Sutterwala SS,Doms RW,Landau NR,Hoxie JA||Cell (87:745)||1996|