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    rankl-antibody-ab9957.pdf

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Immunology Innate Immunity Cytokines TNF Superfamily
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Anti-RANKL antibody (ab9957)

  • Datasheet
  • SDS
Reviews (4)Q&A (3)References (45)

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Abpromise

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Find out more.

Western blot - Anti-RANKL antibody (ab9957)
  • Sandwich ELISA - Anti-RANKL antibody (ab9957)

Key features and details

  • Rabbit polyclonal to RANKL
  • Suitable for: Sandwich ELISA, WB
  • Reacts with: Human
  • Isotype: unknown

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Recombinant mouse RANKL protein (Active) (ab129136)
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Overview

  • Product name

    Anti-RANKL antibody
    See all RANKL primary antibodies
  • Description

    Rabbit polyclonal to RANKL
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Sandwich ELISA, WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Highly pure (>98%) recombinant hsRANK-L (human soluble receptor activator of NF-Kappa B Ligand)

  • General notes

    Although some customers have been successful in IHC we no longer batch test in this application.

    We have received both positive and negative customer feedback on mouse reactivity for this antibody. Therefore, we do not guarantee this species.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

  • Form

    Lyophilized:Reconstitute with 200µl of sterile water. Please note that if you receive this product in liquid form it has already been reconstituted as described and no further reconstitution is necessary.
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
  • Storage buffer

    No preservative, sterile filtered
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    unknown
  • Light chain type

    unknown
  • Research areas

    • Immunology
    • Innate Immunity
    • Cytokines
    • TNF Superfamily
    • Cardiovascular
    • Atherosclerosis
    • Thrombosis
    • Other
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Positive Controls

    • Recombinant human RANKL protein (Active) (ab9958)
  • Recombinant Protein

    • Recombinant mouse RANKL protein (Active) (ab129136)

Applications

Our Abpromise guarantee covers the use of ab9957 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

To detect hsRANKL by sandwich ELISA (using 100 μl/well antibody solution) a concentration of 0.5 - 2.0 μg/ml of this antibody is required. This antigen affinity purified antibody, in conjunction with ab83145 as a detection antibody, allows the detection of at least 0.2 - 0.4 ng/well of recombinant hsRANKL.

WB Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 35 kDa). To detect hsRANK-L by Western Blot analysis this antibody can be used at a concentration of 0.1 - 0.2 µg/ml. Used in conjunction with compatible secondary reagents the detection limit for recombinant hsRANK-L is 1.5 - 3.0 ng/lane, under either reducing or non-reducing conditions.

Target

  • 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.
  • Tissue specificity

    Highest in the peripheral lymph nodes, weak in spleen, peripheral blood Leukocytes, bone marrow, heart, placenta, skeletal muscle, stomach and thyroid.
  • Involvement in disease

    Defects in TNFSF11 are the cause of osteopetrosis autosomal recessive type 2 (OPTB2) [MIM:259710]; also known as osteoclast-poor osteopetrosis. Osteopetrosis is a rare genetic disease characterized by abnormally dense bone, due to defective resorption of immature bone. The disorder occurs in two forms: a severe autosomal recessive form occurring in utero, infancy, or childhood, and a benign autosomal dominant form occurring in adolescence or adulthood. Autosomal recessive osteopetrosis is usually associated with normal or elevated amount of non-functional osteoclasts. OPTB2 is characterized by paucity of osteoclasts, suggesting a molecular defect in osteoclast development.
  • Sequence similarities

    Belongs to the tumor necrosis factor family.
  • Post-translational
    modifications

    The soluble form of isoform 1 derives from the membrane form by proteolytic processing (By similarity). The cleavage may be catalyzed by ADAM17.
  • Cellular localization

    Cytoplasm; Secreted and Cell membrane.
  • Target information above from: UniProt accession O14788 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 8600 Human
    • Omim: 602642 Human
    • SwissProt: O14788 Human
    • Unigene: 333791 Human
    • Alternative names

      • CD254 antibody
      • hRANKL2 antibody
      • ODF antibody
      • OPGL antibody
      • OPTB2 antibody
      • Osteoclast differentiation factor antibody
      • Osteoprotegerin ligand antibody
      • Rank Ligand antibody
      • RANKL antibody
      • Receptor activator of nuclear factor kappa B ligand antibody
      • Receptor activator of nuclear factor kappa-B ligand antibody
      • sOdf antibody
      • TNF related activation induced cytokine antibody
      • TNF-related activation-induced cytokine antibody
      • TNF11_HUMAN antibody
      • TNFSF 11 antibody
      • Tnfsf11 antibody
      • TRANCE antibody
      • Tumor necrosis factor (ligand) superfamily member 11 antibody
      • Tumor necrosis factor ligand superfamily member 11 antibody
      • Tumor necrosis factor ligand superfamily member 11, soluble form antibody
      see all

    Images

    • Western blot - Anti-RANKL antibody (ab9957)
      Western blot - Anti-RANKL antibody (ab9957)
      Anti-RANKL antibody (ab9957) at 1 µg/ml + Human spleen tissue lysate - total protein (ab29699) at 10 µg

      Secondary
      Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed at 1/3000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 35 kDa
      Observed band size: 37 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 72 kDa, 76 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 90 seconds


      RANKL contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
    • Sandwich ELISA - Anti-RANKL antibody (ab9957)
      Sandwich ELISA - Anti-RANKL antibody (ab9957)

      To detect human sRANKL by sandwich ELISA (using 100 μl/well antibody solution) a concentration of 0.25 – 1.0 μg/ml of ab83145 is required. This biotinylated polyclonal antibody, in conjunction with an anti-human sRANKL (ab9957) as a capture antibody, allows the detection of at least 0.2 – 0.4 ng/well of recombinant human sRANKL.

    Protocols

    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (45)

    Publishing research using ab9957? Please let us know so that we can cite the reference in this datasheet.

    ab9957 has been referenced in 45 publications.

    • Wang W  et al. Resveratrol promotes osteoblastic differentiation in a rat model of postmenopausal osteoporosis by regulating autophagy. Nutr Metab (Lond) 17:29 (2020). PubMed: 32322287
    • Maeda T  et al. RANK-RANKL signalling pathway contributes to disease progression in cutaneous angiosarcoma: a case report with an immunohistochemical review and in vitro experiments. J Eur Acad Dermatol Venereol N/A:N/A (2020). PubMed: 32511804
    • Gibo T  et al. Immunohistochemical Investigation of Predictive Biomarkers for Mandibular Bone Invasion in Oral Squamous Cell Carcinoma. Pathol Oncol Res N/A:N/A (2020). PubMed: 32524367
    • Yan K  et al. A20 inhibits osteoclastogenesis via TRAF6-dependent autophagy in human periodontal ligament cells under hypoxia. Cell Prolif 53:e12778 (2020). PubMed: 32027437
    • Zhang X  et al. Knockdown of DANCR reduces osteoclastogenesis and root resorption induced by compression force via Jagged1. Cell Cycle 18:1759-1769 (2019). PubMed: 31213122
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-7 of 7 Abreviews or Q&A

    ELISA abreview for Anti-RANKL antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    ELISA
    Sample
    Human Serum (human serum (serum samples))
    Specification
    human serum (serum samples)
    Type
    Sandwich (Capture)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 09 2013

    Western blot abreview for Anti-RANKL antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    15 µg
    Gel Running Conditions
    Reduced Non-Denaturing (Native) (5-16% gels)
    Sample
    Human Cell lysate - whole cell (human PBMCs)
    Specification
    human PBMCs
    Treatment
    see below (Additional notes)
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 03 2013

    Immunocytochemistry/ Immunofluorescence abreview for Anti-RANKL antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
    Sample
    Human Cell (PBMC³s)
    Specification
    PBMC³s
    Permeabilization
    Yes - 0,1%Saponin/PBS
    Fixative
    Methanol
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 28 2013

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate buffer pH 6.0
    Sample
    Human Tissue sections (bone marrow)
    Specification
    bone marrow
    Permeabilization
    Yes - 0,1% Saponin in PBS
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 05 2013

    Question

    Dear Sirs,
    I'm looking for an anti-human RANKL antibody to be used for WB and immunoistochemistry. I would need to recognize either the soluble and the membrane forms of the protein.
    I have been checking your site but, up to now, I haven't found anything corresponding to all this requisites.
    Could you please give me some help?
    Thank you in advance

    Read More

    Abcam community

    Verified customer

    Asked on Sep 11 2012

    Answer

    Thank you for contacting us.

    We have several antibodies against RANKL.

    Ab9957 is raised against our Recombinant Human sRANKL, which is the soluble form of the protein. However, the listed membrane form of the protein does also contain the soluble portion. As long as the membrane form contains the soluble form and the epitopes are available, the antibody should detect it.

    Regarding ab124797, the sequence used to raise it shares 100% homology with the membrane, cytoplasmic and secreted forms, so it is very likely it will recognise them.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Sep 11 2012

    Question

      Thank you very much.   I am intrested in the antibodies listed below: ab12008 Anti-RANK antibody [9A725] ab 66237 Anti-Cathepsin K ab-49507 Anti-Tartrate Resistant Acid Phosphatase antibody [26E5] - Aminoterminal end As I understand, you will give us some trial free samples for testing, is that right? If yes, I will forward your email to our manager who is taking care of the shipping and receiving materials.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 09 2011

    Answer

    Thank you for your reply. As I mentioned previously, we do not give free samples of our antibodies, even for testing purposes. We issue testing discount codes for antibodies that have not been tested in a specific application or species. How the testing program works is, we would issue you a testing discount code for whichever antibody you choose to test. You would then purchase the antibody at full price, test it in whichever untested application/species you are interested in. You would then submit an Abreview on how the antibody functioned (positive or negative). Once the Abreview has been received, the testing discount that was originally sent to you would become active and this would allow you to receive one antibody free of charge from out catalogue. More information can be found at the link below: https://www.abcam.com/index.htmlpageconfig=resource&rid=11998&viapagetrap=collaborationdiscount For the list of antibodies that you sent me, could you please let me know the application or species you would like to test these antibodies in and I will let you know if they are eligible for a testing discount. I look forward to your reply.    

    Read More

    Abcam Scientific Support

    Answered on Dec 09 2011

    Question

      Thanks for your apply. I am just wondering whether you can offer a small trial amount for each antibody which we can purchase.   For the testing discount program, would you please see whether you have antibodies listed below to try with? Anti-RANK Anti-Cathepsin K Anti-Tartrate Resistant Acid Phosphatase antibody [26E5] –   I will order one vial for RANKL. Thank you.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 07 2011

    Answer

    Thank you for your reply. As mentioned in the previous emails, we do not send samples sizes of our antibodies, under our Abpomise we guarantees the antibodies to work as stated on the datasheet. If the antibodies do not work as stated then we will either replace or refund the antibodies. We give out testing discounts for antibodies that have not been tested in either a specific species or application. Below is a few of the antibodies to your targets: ab12008 Anti-RANK antibody [9A725]   ab13918 Anti-RANK antibody [64C1385   ab19027 Anti-Cathepsin K antibody   ab66237 Anti-Cathepsin K antibody   ab49507 Anti-Tartrate Resistant Acid Phosphatase antibody [26E5] - Aminoterminal end   ab96372 Anti-Tartrate Resistant Acid Phosphatase antibody If none of these antibodies meet your requirements, please let me know which application or species you would like to test them in and I will happily send you a testing discount code. If there is anything else I can help you with, please let me know.  

    Read More

    Abcam Scientific Support

    Answered on Dec 07 2011

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