Anti-CD254 Antibody

Western blot analysis of CD254 expression in COS7 (A), Hela (B), U937 (C), HL60 (D), Raji (E), Ramos (F), Jurkat (G), SW480 (H) whole cell lysates. (Predicted band size: 36 kD; Observed band size: 38 kD)
Immunohistochemical analysis of CD254 staining in human bladder cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
Immunofluorescent analysis of CD254 staining in Hela cells. 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 the primary antibody in 3% BSA-PBS and incubated overnight at 4 °C in a hidified chamber. Cells were washed with PBST and incubated with an AF488-conjugated secondary antibody (green) in PBS at room temperature in the dark. Phalloidin - AF594 was used to stain Actin filaments (red). DAPI was used to stain the cell nuclei (blue).
Flow cytometric analysis of HeLa cells using Anti-CD254 Antibody (green) and negative control (red).

Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC, FC, IF/IC
Primary Accession	O14788
Reactivity	Human, Mk
Host	Mouse
Clonality	Monoclonal
Calculated MW	35478 Da

Additional Information
Gene ID	8600
Other Names	OPGL; RANKL; TRANCE; Tumor necrosis factor ligand superfamily member 11; Osteoclast differentiation factor; ODF; Osteoprotegerin ligand; OPGL; Receptor activator of nuclear factor kappa-B ligand; RANKL; TNF-related activation-induced cytokine; TRANCE; CD254
Target/Specificity	Recombinant fusion protein of human CD254 expressed in E. Coli
Dilution	WB~~WB (1/500 - 1/1000) IHC~~1:100~500 FC~~FC (1/100 - 1/200) IF/IC~~N/A
Format	Mouse IgG1. Liquid in PBS, pH 7.3, 30% glycerol, and 0.01% sodium azide.
Storage	Store at -20 °C.Stable for 12 months from date of receipt

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	TNFSF11
Synonyms	OPGL, RANKL, TRANCE
Function	Cytokine that binds to TNFRSF11B/OPG and to TNFRSF11A/RANK. Osteoclast differentiation and activation factor. Augments the ability of dendritic cells to stimulate naive T-cell proliferation. May be an important regulator of interactions between T-cells and dendritic cells and may play a role in the regulation of the T-cell-dependent immune response. May also play an important role in enhanced bone-resorption in humoral hypercalcemia of malignancy (PubMed:22664871). Induces osteoclastogenesis by activating multiple signaling pathways in osteoclast precursor cells, chief among which is induction of long lasting oscillations in the intracellular concentration of Ca (2+) resulting in the activation of NFATC1, which translocates to the nucleus and induces osteoclast-specific gene transcription to allow differentiation of osteoclasts. During osteoclast differentiation, in a TMEM64 and ATP2A2-dependent manner induces activation of CREB1 and mitochondrial ROS generation necessary for proper osteoclast generation (By similarity).
Cellular Location	[Isoform 1]: Cell membrane; Single-pass type II membrane protein [Isoform 2]: Cytoplasm.
Tissue Location	Highest in the peripheral lymph nodes, weak in spleen, peripheral blood Leukocytes, bone marrow, heart, placenta, skeletal muscle, stomach and thyroid

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.