Anti-RANKL/CD254 antibody (bs-20647R)

Anti-RANKL/CD254 antibody

DATASHEET

Host:Rabbit

Target Protein:RANKL/CD254

IR:Immunogen Range:131-230/317

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:8600

Swiss Prot:O14788

Source:KLH conjugated synthetic peptide derived from human RANKL/CD254:131-230/317 

Purification:affinity purified by Protein A

Storage:0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background:This gene encodes a member of the tumor necrosis factor (TNF) cytokine family which is a ligand for osteoprotegerin and functions as a key factor for osteoclast differentiation and activation. This protein was shown to be a dentritic cell survival factor and is involved in the regulation of T cell-dependent immune response. T cell activation was reported to induce expression of this gene and lead to an increase of osteoclastogenesis and bone loss. This protein was shown to activate antiapoptotic kinase AKT/PKB through a signaling complex involving SRC kinase and tumor necrosis factor receptor-associated factor (TRAF) 6, which indicated this protein may have a role in the regulation of cell apoptosis. Targeted disruption of the related gene in mice led to severe osteopetrosis and a lack of osteoclasts. The deficient mice exhibited defects in early differentiation of T and B lymphocytes, and failed to form lobulo-alveolar mammary structures during pregnancy. Two alternatively spliced transcript variants have been found. [provided by RefSeq, Jul 2008].

Size:100ul

Concentration:1mg/ml

Applications:WB(1:500-2000)
Flow-Cyt(1ug/Test)

Cross Reactive Species:Human
Mouse
Rat
Dog
Horse
Rabbit
.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Sample:
Lymph node(Mouse) Lysate at 40 ug
Lymph node(Rat) Lysate at 40 ug
Primary: Anti-RANKL’CD254 (bs-20647R) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 37’50 kD
Observed band size: 50 kD

Sample:
U937(Human) Cell Lysate at 40 ug
HUVEC(Human) Cell Lysate at 40 ug
Primary: Anti-RANKL/CD254 (bs-20647R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 35 kD
Observed band size: 35 kD

Blank control:HL-60.
Primary Antibody (green line): Rabbit Anti-RANKL/CD254 antibody (bs-20647R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF488
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.