TNFSF11/RANKL Monoclonal antibody

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  WB analysis using 66610-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66610-1-Ig (RANKL antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  WB analysis of HeLa using 66610-1-Ig

  WB result of RANKL antibody (66610-1-Ig; 1:6000; incubated at room temperature for 1.5 hours) with sh-Control and sh-RANKL transfected HeLa cells.

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  WB analysis of DC2.4 using 66610-1-Ig

  DC2.4 cells were subjected to SDS PAGE followed by western blot with 66610-1-Ig (RANKL antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  WB analysis of human spleen using 66610-1-Ig

  human spleen tissue were subjected to SDS PAGE followed by western blot with 66610-1-Ig (RANKL antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 66610-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66610-1-Ig (RANKL antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  WB analysis of U2OS using 66610-1-Ig

  U2OS cells were subjected to SDS PAGE followed by western blot with 66610-1-Ig (RANKL antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  WB analysis of HUVEC using 66610-1-Ig

  HUVEC cells were subjected to SDS PAGE followed by western blot with 66610-1-Ig (RANKL antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  WB analysis of NCCIT using 66610-1-Ig

  NCCIT cell were subjected to SDS PAGE followed by western blot with 66610-1-Ig (RANKL antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

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  IF Staining of MCF-7 using 66610-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using RANKL antibody (66610-1-Ig, Clone: 3F2E1 ) at dilution of 1:800 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

• Featured Product
• KD/KO Validated

TNFSF11/RANKL Monoclonal Antibody for WB, IF/ICC, ELISA

Cat No. 66610-1-Ig

Clone No.3F2E1

Publications(11)

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, ELISA

Synonyms

TNFSF11, 3F2E1, CD 254, CD254, Receptor activator of nuclear factor kappa-B ligand

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Tested Applications

• Positive WB detected in: COLO 320 cells, HeLa cells, U2OS cells, HUVEC cells, NCCIT cells, human spleen tissue, DC2.4 cells, JAR cells, RAW 264.7 cells
• Positive IF/ICC detected in: MCF-7 cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600

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.

Published Applications

• WB: See 9 publications below
• IHC: See 4 publications below
• IF: See 4 publications below

Product Information

66610-1-Ig targets TNFSF11/RANKL in WB, IHC, IF/ICC, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag19975

  Product name: Recombinant human RANKL protein

  Source: e coli.-derived, PET28a

  Tag: 6*His

  Domain: 76-317 aa of BC074890

  Sequence: PNRISEDGTHCIYRILRLHENADFQDTTLESQDTKLIPDSCRRIKQAFQGAVQKELQHIVGSQHIRAEKAMVDGSWLDLAKRSKLEAQPFAHLTINATDIPSGSHKVSLSSWYHDRGWAKISNMTFSNGKLIVNQDGFYYLYANICFRHHETSGDLATEYLQLMVYVTKTSIKIPSSHTLMKGGSTKYWSGNSEFHFYSINVGGFFKLRSGEEISIEVSNPSLLDPDQDATYFGAFKVRDID

• Full Name: tumor necrosis factor (ligand) superfamily, member 11
• Calculated Molecular Weight: 317 aa, 35 kDa
• Observed Molecular Weight: 35-38 kDa
• GenBank Accession Number: BC074890
• Gene Symbol: RANKL
• Gene ID (NCBI): 8600
• RRID: AB_2881970
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G 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.

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
mouse	IHC	J Cancer; Gut microbiota deficiency ameliorates multiple myeloma and myeloma-related bone disease by Th17 cells in mice models; Authors - Jia Liu
rat	WB,IF	Folia Histochem Cytobiol; Platelet-rich plasma ameliorates cartilage degradation in rat models of osteoarthritis via the OPG/RANKL/RANK system; Authors - Qun Wu
human	WB,IF	Cell Death Dis; RANK promotes colorectal cancer migration and invasion by activating the Ca2+-calcineurin/NFATC1-ACP5 axis.; Authors - Qian Liang
mouse	WB,IHC,IF	Open Life Sci; Effect of periostin on bone metabolic and autophagy factors during tooth eruption in mice; Authors - Han Qin
mouse	IF	Adv Healthc Mater; Structured Polymers Enable the Sustained Delivery of Glucocorticoids within the Intra-Articular Space; Authors - Ana Crastin
	WB	Poult Sci; Zinc Glycine supplementation improves bone quality in meat geese by modulating gut microbiota, SCFA's, and gut barrier function through Wnt10b/NF-κB axis; Authors - Zeshan Zulfiqar