Anti-RANKL antibody (ab9957)
- Datasheet
- References (21)
- 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:
- Tang R et al. Interleukin-37 inhibits osteoclastogenesis and alleviates inflammatory bone destruction. J Cell Physiol 234:7645-7658 (2019). Read more (PubMed: 30414292) »
- Deng A et al. The inhibitory roles of Ihh downregulation on chondrocyte growth and differentiation. Exp Ther Med 15:789-794 (2018). Read more (PubMed: 29434683) »
