Anti-RANKL antibody [12A668] - BSA and Azide free (ab45039)

Overview

  • Product name
    Anti-RANKL antibody [12A668] - BSA and Azide free
    See all RANKL primary antibodies
  • Description
    Mouse monoclonal [12A668] to RANKL - BSA and Azide free
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Pmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Bacterially expressed fusion protein, corresponding to amino acids 1-317 of Mouse RANKL

  • Positive control
    • IHC-P: Human lymph node and liver tissue. ICC/IF: HeLa (human epithelial cell line from cervix adenocarcinoma) cells. WB: Human lymph node lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab45039 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
ICC/IF Use a concentration of 10 µg/ml.
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 35 kDa (predicted molecular weight: 35 kDa).
IHC-P Use a concentration of 1 - 5 µg/ml.
  • Application notes
    Is unsuitable for Flow Cyt.
  • 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.
    • 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
      • 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 [12A668] - BSA and Azide free images

    • Paraffin embedded human lymph node tissue stained for RANKL with ab45039 at 5 µg/ml.

    • ICC/IF image of ab45039 stained HeLa cells. The cells were 4% formaldehyde fixed (10 minutes) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab45039, 10 µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.

    • Anti-RANKL antibody [12A668] - BSA and Azide free (ab45039) at 2 µg + transfected cell lysate

      Predicted band size : 35 kDa
      Observed band size : 35 kDa
    • Paraffin embedded human liver tissue stained for RANKL with ab45039 at a 1/500 dilution. Peroxidase-conjugate and DAB chromogen. A 2 hour incubation at RT was used.

    • Anti-RANKL antibody [12A668] - BSA and Azide free (ab45039) at 0.5 µg/ml + human lymph node lysate in RIPA buffer at 35 µg

      Predicted band size : 35 kDa
      Additional bands at : 28 kDa (possible isoform).

      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)

    References for Anti-RANKL antibody [12A668] - BSA and Azide free (ab45039)

    This product has been referenced in:
    • Xu Z  et al. SMURF2 regulates bone homeostasis by disrupting SMAD3 interaction with vitamin D receptor in osteoblasts. Nat Commun 8:14570 (2017). Read more (PubMed: 28216630) »
    • Becerikli M  et al. Age-dependent alterations in osteoblast and osteoclast activity in human cancellous bone. J Cell Mol Med N/A:N/A (2017). IHC-P ; Human . Read more (PubMed: 28444839) »

    See all 9 Publications for this product

    Product Wall

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (mandible)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Buffer solutions (Citrate pH 6,0)
    Permeabilization
    No
    Specification
    mandible
    Blocking step
    Hydrogen Peroxide as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Aug 30 2016

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Western blot
    Sample
    Rabbit Tissue lysate - whole (Subchondral bone)
    Gel Running Conditions
    Reduced Denaturing (Gel 15%)
    Loading amount
    30 µg
    Specification
    Subchondral bone
    Blocking step
    Milk 5%+ BSA 3% as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Username

    Abcam user community

    Verified customer

    Submitted Jul 01 2016

    Thank you for your enquiry.

    All tested applications and species covered by our 6 month guarantee are specified on our datasheets, and these are updated as soon as any new information is brought to our attention.

    Therefore, I can c...

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    I have received confirmation that these 3 replacement vials will be included in the next shipment to Aurogene. I hope this is helpful. Please contact me again if you have any further questions.

    Thanks for your reply. It seems that, as part of their own internal QC, the customer has already peformed the control I suggested. This is great as it does appear clear now that these 3 antibodies are not working as expected. I would be happy to reques...

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    Thank you for your reply. I am pleased to provide the following information: We have two RANK antibodies tested flow cytometry, human and mouse (not IHC or rat): ab45092 RANK antibody [9A725] (Phycoerythrin) Mouse monoclonal Tested applicat...

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    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody. The details you have kindly provided will enable us to investigate this case f...

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    Thank you for providing the additional information. The one thing I would like to recommend in this case is to run a "loading control" western blot with the murine osteoblast cell lysate. Examples of targets for this type of control would be proteins t...

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    Thank you for your enquiry. Sorry to hear your customer's are having trouble with these antibodies.   What species of bone cells were being tested? What dilution of primary antibody for each? Was any other antibody used successfully with these same sam...

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    Thank you for your enquiry regarding ab45039 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody. I can confirm that anti RANKL antibody is indeed specif...

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