TNFSF11/RANKL Recombinant monoclonal antibody

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• TNFSF11/RANKL Recombinant monoclonal antibody (85321-2-RR)

TNFSF11/RANKL Uni-rAb^® Recombinant Antibody for WB, IF/ICC, FC (Intra), ELISA

Cat No. 85321-2-RR

Clone No.251930C5

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

WB, IF/ICC, FC (Intra), ELISA

Synonyms

TNFSF11, CD 254, CD254, Receptor activator of nuclear factor kappa-B ligand, TNF-related activation-induced cytokine

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  WB analysis using 85321-2-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 85321-2-RR (TNFSF11/RANKL antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  WB analysis of Raji using 85321-2-RR

  Raji cells were subjected to SDS PAGE followed by western blot with 85321-2-RR (TNFSF11/RANKL antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  IF Staining of THP-1 using 85321-2-RR

  Immunofluorescent analysis of (4% PFA) fixed THP-1 cells using TNFSF11/RANKL antibody (85321-2-RR, Clone: 251930C5 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2).

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  FC experiment of MJ using 85321-2-RR

  1x10^6 MJ cells were intracellularly stained with 0.25 ug TNFSF11/RANKL Recombinant monoclonal antibody (85321-2-RR, Clone:251930C5) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.25 ug Rabbit IgG Isotype Control RecAb (98136-1-RR, Clone: 240953C9) (blue). Cells were fixed and permeabilized with Intracellular Flow Cytometry Fixation & Permeabilization Buffer Kit (PF00019).

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  Affinity and Kinetic Characterization of 85321-2-RR

  Biolayer interferometry (BLl) kinetic assays of 85321-2-RR against Human TNFSF11/RANKL were performed. The affinity constant is 5.52 nM.

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Tested Applications

• Positive WB detected in: THP-1 cells, Raji cells, RAW 264.7 cells
• Positive IF/ICC detected in: THP-1 cells
• Positive FC (Intra) detected in: MJ cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

85321-2-RR targets TNFSF11/RANKL in WB, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen:

  CatNo: Eg2564

  Product name: Recombinant Human TNFSF11/RANKL protein (rFc Tag)

  Source: mammalian cells-derived, pHZ-KIsec-N-rFc

  Tag: N-rFc

  Domain: 64-245 aa of NM_003701

  Sequence: GSQHIRAEKAMVDGSWLDLAKRSKLEAQPFAHLTINATDIPSGSHKVSLSSWYHDRGWGKISNMTFSNGKLIVNQDGFYYLYANICFRHHETSGDLATEYLQLMVYVTKTSIKIPSSHTLMKGGSTKYWSGNSEFHFYSINVGGFFKLRSGEEISIEVSNPSLLDPDQDATYFGAFKVRDID

• Full Name: tumor necrosis factor (ligand) superfamily, member 11
• Calculated Molecular Weight: 28KD
• Observed Molecular Weight: 35 kDa
• GenBank Accession Number: NM_003701
• Gene Symbol: RANKL
• Gene ID (NCBI): 8600
• RRID: AB_3744134
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• UNIPROT ID: O14788
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

TNFSF11 also known as RANKL, is 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. RANKL is a polypeptide of 217 amino acids that exerts its biological activity both in a transmembrane form of about 40-45 kDa and in soluble one of 31 kDa (PMID: 15308315). The membrane-bound RANKL (mRANKL) is cleaved into a sRANKL by the metalloprotease-disintegrin TNF-alpha convertase (TACE) or a related metalloprotease (MP). RANKL induces osteoclast formation through its receptor, RANK, which transduces signals by recruiting adaptor molecules, such as the TNF receptor-associated factor (TRAF) family of proteins. RANKL 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. RANKL 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.