TRANCE/TNFSF11/RANK L Antibody (12A668) - BSA Free

Immunohistochemistry: TRANCE/TNFSF11/RANK L Antibody (12A668) [NB100-56512] - MQDL detects EMT biomarkers in clinical prostate cancer. EMT biomarkers (EpCAM, N-Cad, and RANKL)were detected in a clinical primary prostate cancer specimen (Gleason score 6; 3+3) with documented bone metastasis. M denotes cells that completed EMT and E/M indicates cells undergoing partial EMT. 400x. Image collected and cropped by CiteAb from the following publication (//journals.plos.org/plosone/article?id=10.1371/journal.pone.0028670) licensed under a CC-BY license.

Western Blot: TRANCE/TNFSF11/RANK L Antibody (12A668) [NB100-56512] - Analysis using Azide Free version of NB100-56512. Human lymph node lysate (35ug per lane, RIPA buffer). Band detected at ~35kDa and ~28kDa. (Expected MW of 35.5kDa according to NP_003692.1 and of 27.7kDa according to NP_143026.1)

Immunocytochemistry/Immunofluorescence: TRANCE/TNFSF11/RANK L Antibody (12A668) [NB100-56512] - TRANCE was detected in immersion fixed mouse splenocytes using anti-human/mouse/rat mouse monoclonal antibody (Catalog # NB100-56512) for 1 hour at room temperature. Cells were stained using NL557 (red) fluorescent anti-mouse secondary antibodies (Catalog # NL007) and counterstained with DAPI (blue).

Immunohistochemistry: TRANCE/TNFSF11/RANK L Antibody (12A668) [NB100-56512] - MQDL detects EMT biomarkers in clinical bone tissue specimens. A representative specimen of human prostate cancer bone metastasis co-expressed high levels of epithelial EpCAM, and mesenchymal RANKL and vimentin proteins. 400x. Image collected and cropped by CiteAb from the following publication (//journals.plos.org/plosone/article?id=10.1371/journal.pone.0028670) licensed under a CC-BY license.

Western Blot: TRANCE/TNFSF11/RANK L Antibody (12A668) [NB100-56512] - Analysis of transfected cell lysate was probed with TRANCE/TNFSF11/RANK L antibody.

Immunohistochemistry-Paraffin: TRANCE/TNFSF11/RANK L Antibody (12A668) [NB100-56512] - Analysis using Azide Free version of NB100-56512. FFPE human liver stained with TRANCE/TNFSF11/RANK L antibody, peroxidase-conjugate and DAB chromogen. A 2 hr incubation at RT was used.

Immunohistochemistry-Paraffin: TRANCE/TNFSF11/RANK L Antibody (12A668) [NB100-56512] - Analysis using Azide Free version of NB100-56512. FFPE human lymph node probed with TRANCE/TNFSF11/RANK L antibody.

MQDL detects EMT biomarkers in clinical bone tissue specimens.A representative specimen of human prostate cancer bone metastasis co-expressed high levels of epithelial EpCAM, and mesenchymal RANKL and vimentin proteins. ×400.

Immunocytochemistry/ Immunofluorescence: TRANCE/TNFSF11/RANK L Antibody (12A668) - BSA Free [NB100-56512] - MQDL detects EMT biomarkers in clinical bone tissue specimens.A representative specimen of human prostate cancer bone metastasis co-expressed high levels of epithelial EpCAM, & mesenchymal RANKL & vimentin proteins. ×400. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0028670), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ELISA, Flow, ICC/IF, IHC, ChIP
• Clone: 12A668
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

TRANCE/TNFSF11/RANK L Antibody (12A668) - BSA Free Summary

• Immunogen: A bacterially expressed fusion protein containing amino acid residues 1-317 of mouse TRANCE/TNFSF11/RANK L was used as immunogen.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: TNFSF11
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 1:10-1:500
  • ELISA reported in scientific literature (PMID 15935726)
  • Flow (Cell Surface)
  • Immunoassay
  • Immunocytochemistry/ Immunofluorescence 20 ug/ml
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Paraffin 5 ug/ml
  • Western Blot 0.5-2 ug/ml
• Theoretical MW: 35 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  TRANCE/TNFSF11/RANK L Overexpression Lysate

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein G purified

Alternate Names for TRANCE/TNFSF11/RANK L Antibody (12A668) - BSA Free

• CD254 antigen
• CD254
• ODF
• OPGL
• OPGLOPTB2
• Osteoclast differentiation factor
• Osteoprotegerin ligand
• RANK L
• RANKL
• RANKLreceptor activator of nuclear factor kappa B ligand
• Receptor activator of nuclear factor kappa-B ligand
• sOdf
• TNF-related activation-induced cytokine
• TNFSF11
• TRANCE
• TRANCEODFhRANKL2
• tumor necrosis factor (ligand) superfamily, member 11
• tumor necrosis factor ligand superfamily member 11

Background

TRANCE/OPGL/RANKL/ODF is a recently identified member of tumor necrosis factor family. TRANCE (TNF-related activation-induced cytokine receptor) or RANKL {receptor activator of NF-kB (RANK) ligand} has been implicated in interactions between T cells and dendritic cells. RANK ligand (RANKL/TRANCE) binds to RANK on dendritic cells, upregulates the expression of anti-apoptotic protein BcL-XL suggesting a role in dendritic cell survival. TRANCE/RANKL is also important in T and B-cell maturation. As OPGL/ODF (Osteoprotegerin Ligand/Osteoclast differentiation factor), the same protein can both activate mature osteoclasts and mediate osteoclastogenesis. OPGL/TRANCE deficient mice show severe osteoporesis and complete absence of osteoclasts as a result of lack of osteogenesis.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for TRANCE/TNFSF11/RANK L Antibody (NB100-56512)(38)

Publications using NB100-56512

• Fei Wei, Megan Hughes, Mahmoud Omer, Christopher Ngo, Abinaya Sindu Pugazhendhi, Elayaraja Kolanthai, Matthew Aceto, Yasmine Ghattas, Mehdi Razavi, Thomas J Kean, Sudipta Seal, Melanie Coathup A Multifunctional Therapeutic Strategy Using P7C3 as A Countermeasure Against Bone Loss and Fragility in An Ovariectomized Rat Model of Postmenopausal Osteoporosis. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2024-03-13 [PMID: 38477537]
• Odo A, Kunimatsu R, Abe T et al. Stem cells derived from human exfoliated deciduous teeth-based media in a rat root resorption model Archives of Oral Biology 2023-11-01 [PMID: 38056228] (IHC, Rat)
• Hild V, Mellert K, Möller P, Barth TFE Giant Cells of Various Lesions Are Characterised by Different Expression Patterns of HLA-Molecules and Molecules Involved in the Cell Cycle, Bone Metabolism, and Lineage Affiliation: An Immunohistochemical Study with a Review of the Literature Cancers 2023-07-21 [PMID: 37509363] (Immunohistochemistry-Paraffin, Human)
  
  Details:
  1:400 IHC-P dilution
• Wei F, Tuong ZK, Omer M et al. A novel multifunctional radioprotective strategy using P7C3 as a countermeasure against ionizing radiation-induced bone loss Bone research 2023-06-29 [PMID: 37385982] (IHC-P, Rat)
• Liu J, Yue J, Wang K et al. Tertiary Lymphoid Structures Are Related to Inflammatory Progression and Bone Loss in Human Apical Periodontitis Journal of endodontics 2023-06-17 [PMID: 37331649]
• Kunimatsu R, Kimura A, Sakata S et al. Effects of baicalin on the proliferation and expression of OPG and RANKL in human cementoblast-lineage cells J Dent Sci 2022-01-14 [PMID: 35028034]
• Lysitska A, Galanis N, Skandalos I Et al. Histology and Immunohistochemistry of Radial Arteries Are Suggestive of an Interaction between Calcification and Early Atherosclerotic Lesions in Chronic Kidney Disease Medicina 2021-10-24 [PMID: 34833374] (IHC-P, Human)
• Liu X, Zhu R, Luo Y et al. Distinct human Langerhans cell subsets orchestrate reciprocal functions and require different developmental regulation Immunity 2021-09-08 [PMID: 34508661]
• Leite FRM, de Aquino SG, Guimaraes MR. RANKL expression is differentially modulated by TLR2 and TLR4 signaling in fibroblasts and osteoblasts Immunology Innovation. (WB, Mouse, Rat)
  
  Details:
  RANKL antibody used for WB for detecting the protein expression of receptor activator of nuclear factor-kappa B ligand /RANKL induced by E coli lipopolysaccharide/LPS, Porphyromonas gingivalis LPS and interleukin-1 beta in Mouse periodontal ligament fibroblasts (mPDL; immortalized with simian virus 40 large T antigen) and Osteoblastic rat osteosarcoma cells ROS 17/2.8 (ROS)
• Sengun MC, Gunpinar S Effects of systemic hydroxytyrosol application in experimental periodontitis of rats International Journal of Plant Based PHARMACEUTICALS 2022-01-01 (IF/IHC, Rat)
• Nakao Y, Fukuda T, Zhang Q et al. Exosomes from TNF-alpha-treated human gingiva-derived MSCs enhance M2 macrophage polarization and inhibit periodontal bone loss Acta biomaterialia 2020-12-24 [PMID: 33359765] (WB, Human)
• Kunimatsu R, Kimura A, Tsuka Y et al. Baicalin inhibits root resorption during tooth movement in a rodent model Arch. Oral Biol. 2020-05-19 [PMID: 32470833] (IF/IHC, Rat)
• Hong H, Hosomichi J, Maeda H et Al. Intermittent hypoxia retards mandibular growth and alters RANKL expression in adolescent and juvenile rats Eur J Orthod 2020-03-27 [PMID: 32219305] (Rat)
• Liu X, Li W, Cai J, et al. Spatiotemporal characterization of microdamage accumulation and its targeted remodeling mechanisms in diabetic fatigued bone FASEB j. 2019-12-18 [PMID: 31908007] (IF/IHC, Rat)
• Gegen T, Zhu Y, Sun QHou B Role of interleukin-33 in the clinical pathogenesis of chronic apical periodontitis J. Int. Med. Res. 2019-06-20 [PMID: 31218936] (IF/IHC, Human)
• Tanaka T, Kelly M, Takei Y, Yamanouchi D. RANKL-mediated osteoclastogenic differentiation of macrophages in the abdominal aorta of angiotensin II-infused apolipoprotein E knockout mice. J Vasc Surg 2018-04-20 [PMID: 29685509] (ICC/IF, Mouse)
  
  Details:
  This citation used the Azide Free version of this antibody.
• Gunji H, Kunimatsu R, Tsuka Y, Yoshimi Y. Effect of high-frequency near-infrared diode laser irradiation on periodontal tissues during experimental tooth movement in rats. Lasers Surg Med. 2018-02-05 [PMID: 29399884] (Rat)
• Shao J, Zhang Y, Yang T et al. HIF-1alpha disturbs osteoblasts and osteoclasts coupling in bone remodeling by up-regulating OPG expression In Vitro Cell. Dev. Biol. Anim. 2015-04-10 [PMID: 25859979] (WB, Chemotaxis, Mouse)
• Callegari A, Coons ML, Ricks JL et al. Increased Calcification in Osteoprotegerin-Deficient Smooth Muscle Cells: Dependence on Receptor Activator of NF-kB Ligand and Interleukin 6. J. Vasc. Res. 2014-03-07 [PMID: 24642764] (IHC-P, Mouse)
  
  Details:
  Mouse innominate arteries, Fig 5.
• Kim RY, Oh JH, Lee BS et al. The effect of dose on rhBMP-2 signaling, delivered via collagen sponge, on osteoclast activation and in vivo bone resorption. Biomaterials 2014-02-01 [PMID: 24321706] (IHC-P, Rat)
• Kresnoadi U, Laksono V, Dahlan A Expression and ratio of receptor activator of nuclear factor kappa-B ligand and osteoprotegerin following application of Nigella sativa/bovine bone graft combination in post tooth extraction sockets The Journal of Indian Prosthodontic Society 2023-07-14 (Immunohistochemistry-Paraffin, Guinea Pig)
• Mine Y, Makihira S, Yamaguchi Y et al. Involvement of ERK and p38 MAPK pathways on Interleukin-33-induced RANKL expression in osteoblastic cells. Cell Biol. Int. 2014-01-22 [PMID: 24448827] (WB, Mouse)
  
  Details:
  MC3T3-E1 mouse calvaria cell line, Fig 2B. IL-33 stimulation upregulated RANkL.
• Hu P, Chung LW, Berel D et al. Convergent RANK- and c-Met-Mediated Signaling Components Predict Survival of Patients with Prostate Cancer: An Interracial Comparative Study. PLoS One. 2013-09-16 [PMID: 24066029] (Human)
• Gambichler T, Terras S, Kampilafkos P et al. T regulatory cells and related immunoregulatory factors in polymorphic light eruption following ultraviolet A1 challenge. Br J Dermatol 2013-12-01 [PMID: 24032533] (IHC-P, Human)
• Yin Y, Schluesener HJ, Zhang ZY, Zhang Z. Lesional infiltration of receptor activator of nuclear factor-kB ligand+ cells in experimental autoimmune neuritis rats Neurol Sci 2012-07-19 [PMID: 22810119] (IF/IHC, Rat)
• Boyce AM, Chong WH, Yao J et al. Denosumab treatment for fibrous dysplasia. J Bone Miner Res. 2012-07-01 [PMID: 22431375] (IHC-P, Mouse)
• Hu P, Chu GC, Zhu G et al. Multiplexed quantum dot labeling of activated c-Met signaling in castration-resistant human prostate cancer. PLoS One. 2011-01-01 [PMID: 22205960] (ICC/IF, IF/IHC, Human)
• Shin M, Matsuo K, Tada T et al. The inhibition of RANKL/RANK signaling by osteoprotegerin suppresses bone invasion by oral squamous cell carcinoma cells. Carcinogenesis. 2011-11-01 [PMID: 21890459]
• Parrula C, Zimmerman B, Nadella P et al. Expression of tumor invasion factors determines systemic engraftment and induction of humoral hypercalcemia in a mouse model of adult T-cell leukemia. Vet Pathol. 2009-09-01 [PMID: 19429977] (Flow-CS, Human)
  
  Details:
  RankL (IMG-133A) for Flow (cell surface), Fig 9 (MET-1 human leukemia cells). 33% of the cells were positive for RankL expression.
• Ghoreishi M, Bach P, Obst J et al. Expansion of antigen-specific regulatory T cells with the topical vitamin d analog calcipotriol. J Immunol. 2009-05-15 [PMID: 19414758] (IHC-P, Mouse)
• Neidhart M, Baraliakos X, Seemayer C et al. Expression of cathepsin K and matrix metalloproteinase 1 indicate persistent osteodestructive activity in long-standing ankylosing spondylitis. Ann Rheum Dis. 2009-08-01 [PMID: 18678577] (IHC-P, Human)
  
  Details:
  IHC (paraffin): Fig 6, spinal tissues from human anylosing spondylitis (RANkL positive) and degenerative disk disease (RANkL negative).
• Sandra F, Hendarmin L, Kukita T et al. Ameloblastoma induces osteoclastogenesis: a possible role of ameloblastoma in expanding in the bone. Oral Oncol. 2005-07-01 [PMID: 15935726] (ELISA)
  
  Details:
  Ameloblastoma tumor tissue and AM-1 cells: 1. OPG (IMG-103A), ELISA (detection): Results described.
• Rubin J, Chung LW, Fan X et al. Prostate carcinoma cells that have resided in bone have an upregulated IGF-I axis. Prostate. 2004-01-01 [PMID: 14673951]
• Niida S, Kawahara M, Ishizuka Y et al. Gamma-glutamyltranspeptidase stimulates receptor activator of nuclear factor-kappaB ligand expression independent of its enzymatic activity and serves as a pathological bone-resorbing factor. J Biol Chem. 2004-02-13 [PMID: 14634009]
• Fiumara P, Snell V, Li Y et al. Functional expression of receptor activator of nuclear factor kappaB in Hodgkin disease cell lines. Blood. 2001-11-01 [PMID: 11675352]
• Pearse RN, Sordillo EM, Yaccoby S et al. Multiple myeloma disrupts the TRANCE/ osteoprotegerin cytokine axis to trigger bone destruction and promote tumor progression. Proc Natl Acad Sci U S A. 2001-09-25 [PMID: 11562486]
  
  Details:
  Antibodies cited [IHC (paraffin)], Figs 1 and 4 (human plasmocytoma and decalified bone marrow): 1. OPG (IMG-103) 2. Trance/RankL/OPGL/ODF (IMG-133).
• Kikuchi T, Matsuguchi T, Tsuboi N et al. Gene expression of osteoclast differentiation factor is induced by lipopolysaccharide in mouse osteoblasts via Toll-like receptors. J Immunol. 2001-03-01 [PMID: 11207318]
• Hsu H, Lacey DL, Dunstan CR et al. Tumor necrosis factor receptor family member RANK mediates osteoclast differentiation and activation induced by osteoprotegerin ligand. Proc Natl Acad Sci U S A. [PMID: 10097072]
  
  Details:
  Citation using the Azide Free form of this antibody.

FAQs for TRANCE/TNFSF11/RANK L Antibody (NB100-56512). (Showing 1 - 3 of 3 FAQs).

1. We have received a customer inquiry regarding your RANKL Antibody (12A668) 0.1mg, Cat # NB100-56512 (previously from Imgenex). Specifically, they would like to know if this product is suitable for use in immunoprecipitation, also, what is the level of cross reactivity to mouse compared to the target reactivity?
   • This product has been validated in mouse, so it is 100% guaranteed for use on mouse samples. It has never been directly tested by Novus or Imgenex for IP, but it works in ICC and for surface staining in Flow Cytometry. Because of this, I would expect that it would be suitable for use in IP as well.
2. I need a positive control in order to establish them for IHC-P. Do you have any further recommendations except the ones stated on the website. For example what kind of tissue could be used as a positive control?
   • RANK/TNFRSF11A demonstrates ubiquitous expression with high levels found in skeletal muscle, thymus, liver, colon, small intestine and adrenal gland. You may use any of these tissues as positive controls. For additional options, you may refer to the staining results posted at Human Protein atlas database and in some IHC-P related publications at online product sheet.
3. Does your RANKL antibody (NB100-56512, Clone 12A668) block the ligand binding site? (or what the epitope range is?) Furthermore, can this antibody use as agonistic?
   • This antibody was raised against a bacterially expressed fusion protein containing amino acid residues 1-317 / whole sequence of mouse TRANCE was used as immunogen (NP_003692; uniprot site). We have not epitope mapped this antibody and accordingly, we are not sure where on RANKL does this antibody bind. We are not aware of its use as agonist or antigonist but since this antibody worked really well in Flow, ICC/IF and IHC-P (applications wherein the antibody binds the antigen in more of native state), it is quite possible that it may work for functional application under in-vitro or in-vivo conditions.

Bioinformatics

• Gene Symbol: TNFSF11
• Entrez:
  • 21943(Mouse)
• Uniprot:
  • O14788()