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.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|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.|
|Storage buffer||whole serum|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:1000 - 1:5000|
|Immunocytochemistry (ICC)||1:1000 - 1:5000|
|Immunofluorescence (IF)||1:1000 - 1:5000|
|Immunohistochemistry (Paraffin) (IHC (P))||1:1000 - 1:5000|
|Western Blot (WB)||1:1000 - 1:5000|
* 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.
|Immunohistochemistry (IHC)||See 1 publications below|
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).
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. It acts as a receptor for extracellular ubiquitin; leading to enhanced intracellular calcium ions and reduced cellular cAMP levels. It is involved in hematopoiesis and in cardiac ventricular septum formation. CXCR4 also plays an essential role in vascularization of the gastrointestinal tract, probably by regulating vascular branching and/or remodeling processes in endothelial cells. Involved in cerebellar development. In the CNS, could mediate hippocampal-neuron survival. It acts as a coreceptor (CD4 being the primary receptor) for HIV-1 X4 isolates and as a primary receptor for some HIV-2 isolates, and promotes Env-mediated fusion of the virus. CXCR4 binds bacterial lipopolysaccharide (LPS) et mediates LPS-induced inflammatory response, including TNF secretion by monocytes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
CXCR4 regulates interneuron migration in the developing neocortex.
PA3-305 was used in immunohistochemistry to investigate the important role of CXCR4 in interneuron migration
|Stumm RK,Zhou C,Ara T,Lazarini F,Dubois-Dalcq M,Nagasawa T,Höllt V,Schulz S||The Journal of neuroscience : the official journal of the Society for Neuroscience (23:5123)||2003|