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详细介绍
Overview
Product name
Anti-RANKL antibody
See all RANKL products （13） ...
Description
Rabbit polyclonal to RANKL
Tested applications
ICC/IF, ELISA, WB, IHC-Pmore details
Cross reactivity
Reacts with
Mouse, Human
Immunogen
Highly pure （>98%） recombinant hsRANK-L （human soluble receptor activator of NF-Kappa B Ligand）
Properties
Form
Lyophilised:
Reconstitute with 200µl of sterile water.
Storage instructions
Store at +4°C short term （1-2 weeks）. Aliquot and store at -20°C long term. Avoid repeated freeze / thaw cycles.
Storage buffer
PBS, pH 7.4, no preservative, sterile filtered
Concentration
Reconstitution dependent
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
unknown
Light chain type
unknown
Research areas
Immunology >> Adaptive Immunity >> Regulatory T Cells
Cardiovascular >> Atherosclerosis >> Thrombosis >> Other
Immunology >> Innate Immunity >> Cytokines >> TNF Superfamily
- Datasheet PDF
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Immunohistochemistry （Formalin/PFA-fixed paraffin-embedded sections） - RANKL antibody （ab9957）
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Immunocytochemistry/ Immunofluorescence-RANKL antibody（ab9957）
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Western blot - RANKL antibody （ab9957）
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Applications
Show applications key
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.
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ICC/IF
ICC/IF: Use a concentrat...Read more →ICC/IF: Use a concentration of 1 µg/ml.
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ELISA
ELISA: Use at an assay d...Read more →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.
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WB
WB: Use a concentration ...Read more →WB: Use a concentration of 1 µg/ml. Detects a band of approximay 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.
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IHC-P
IHC-P: Use at an assay d...Read more →IHC-P: Use at an assay dependent dilution.
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
Alternative names
- CD254 antibody
- hRANKL2 antibody
- ODF antibody
see all
Database links
- Entrez Gene: 8600 Human
- Entrez Gene: 21943 Mouse
- Omim: 602642 Human
- SwissProt: O14788 Human
- SwissProt: O35235 Mouse
- Unigene: 333791 Human
- Unigene: 249221 Mouse
Anti-RANKL antibody images:
Immunohistochemistry （Formalin/PFA-fixed paraffin-embedded sections） - 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-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.
Western blot - RANKL antibody （ab9957）
（ab9957） at 1 µg/ml +
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.
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