Anti-RANKL antibody (ab9957)
- Datasheet
- References (19)
- Protocols
Overview
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Product nameAnti-RANKL antibody
See all RANKL primary antibodies -
DescriptionRabbit polyclonal to RANKL
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Host speciesRabbit
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Tested applicationsSuitable for: ICC/IF, ELISA, WB, IHC-Pmore details
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Species reactivityReacts with: Human
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Immunogen
Highly pure (>98%) recombinant hsRANK-L (human soluble receptor activator of NF-Kappa B Ligand)
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General notes
We have received both positive and negative customer feedback on mouse reactivity for this antibody. Therefore, we do not guarantee this species.
Properties
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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.
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
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Storage bufferPBS, pH 7.4, no preservative, sterile filtered
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Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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Isotypeunknown
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Light chain typeunknown
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Research areas
Associated products
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Compatible Secondaries
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Positive Controls
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 |
|---|---|---|
| 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
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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.
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Tissue specificityHighest in the peripheral lymph nodes, weak in spleen, peripheral blood Leukocytes, bone marrow, heart, placenta, skeletal muscle, stomach and thyroid.
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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.
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Sequence similaritiesBelongs to the tumor necrosis factor family.
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Post-translational
modificationsThe 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.
- Information by UniProt
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Database links
- Entrez Gene: 8600 Human
- Omim: 602642 Human
- SwissProt: O14788 Human
- Unigene: 333791 Human
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Alternative names
- CD254 antibody
- hRANKL2 antibody
- ODF antibody
see all
Images
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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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RANKL antibody (ab9957)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. -
Immunocytochemistry/ Immunofluorescence - Anti-RANKL antibody (ab9957)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.
Protocols
References
This product has been referenced in:
- Scotto di Carlo F et al. The identification of H3F3A mutation in giant cell tumour of the clivus and the histological diagnostic algorithm of other clival lesions permit the differential diagnosis in this location. BMC Cancer 18:358 (2018). Read more (PubMed: 29609578) »
- Zhang L et al. Dishevelled-2 modulates osteogenic differentiation of human synovial fibroblasts in osteoarthritis. Mol Med Rep 18:292-298 (2018). Read more (PubMed: 29749449) »
