Overview

  • Product nameAnti-RANKL antibodySee all RANKL primary antibodies ...
  • Description
    Rabbit polyclonal to RANKL
  • Tested applicationsICC/IF, ELISA, WB, IHC-P more details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

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

Properties

  • FormLyophilised: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 instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freeze / thaw cycles.
  • Storage bufferPBS, pH 7.4, no preservative, sterile filtered
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • Clonality Polyclonal
  • Isotypeunknown
  • Light chain typeunknown
  • Research Areas

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 Notes
ICC/IF Use a concentration of 1 µg/ml.
ELISA Use at an assay dependent dilution. To detect hsRANK-L by direct ELISA (using 100µl/well antibody solution) a concentration of at least 0.5µg/ml of this antibody is required. This antigen affinity purified antibody, in conjunction with compatible secondary reagents, allows the detection of 0.2 - 0.4 ng/well of recombinant hsRANK-L.
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.
IHC-P Use at an assay dependent dilution.

Target

  • FunctionCytokine 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 specificityHighest in the peripheral lymph nodes, weak in spleen, peripheral blood Leukocytes, bone marrow, heart, placenta, skeletal muscle, stomach and thyroid.
  • Involvement in diseaseDefects 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 similaritiesBelongs 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 localizationCytoplasm; 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
  • Alternative names
    • CD254 antibody
    • hRANKL2 antibody
    • ODF antibody
    • OPGL antibody
    • OPTB2 antibody
    • Osteoclast differentiation factor antibody
    • Osteoprotegerin ligand antibody
    • RANKL antibody
    • Receptor activator of nuclear factor kappa B ligand antibody
    • Receptor activator of nuclear factor kappa-B ligand antibody
    • sOdf antibody
    • SOFA 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

Anti-RANKL antibody images

  • ab9957 staining RANKL in human metastatic carcinoma of lymph nodes from breast cancer tissue by Immunohistochemistry (Formalin/PFA fixed paraffin-embedded sections). Tissue underwent heat mediated antigen retrieval in sodium citrate buffer (pH 6.0). The primary antibody was used at 0.25 ug/ml and incubated with sample at 4°C overnight. A HRP-labeled polymer detection system was used with a DAB chromogen.
  • ICC/IF image of ab9957 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9957, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Anti-RANKL antibody (ab9957) at 1 µg/ml + Spleen (Human) Tissue Lysate - adult normal tissue (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

References for Anti-RANKL antibody (ab9957)

This product has been referenced in:
  • Peng X  et al. Differential expression of the RANKL/RANK/OPG system is associated with bone metastasis in human non-small cell lung cancer. PLoS One 8:e58361 (2013). WB, IHC-P ; Human . Read more (PubMed: 23516466) »
  • Mediero A  et al. Adenosine A2A receptor activation prevents wear particle-induced osteolysis. Sci Transl Med 4:135ra65 (2012). IHC-P ; Mouse . Read more (PubMed: 22623741) »

See all 3 Publications for this product

Product Wall

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
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Submitted Jul 09 2013

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
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Submitted Jul 03 2013

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
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Submitted Jun 28 2013

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
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Submitted Jun 05 2013

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 conta...

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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 testin...

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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 eith...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"