Antibody data
- Antibody Data
- Antigen structure
- References [2]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [1]
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Validation data
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- Product number
- 14-6619-82 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD254 (RANK Ligand) Monoclonal Antibody (MIH24), eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The MIH24 monoclonal antibody reacts with human TRANCE (TNF-related activation-induced cytokine receptor) also known as RANKL (receptor activator of NF-kappaB ligand) and OPGL (Osteoprotegerin Ligand). TRANCE is a type II membrane protein with predicted molecular weight of 35 kDa. Its extracellular domain is most closely related to TRAIL, FasL and TNF. TRANCE mRNA is upregulated by TCR stimulation. TRANCE-deficient mouse lacks osteoclasts. In addition, TRANCE has been shown to be involved in the interactions between T-dendritic and T-B cells. It was also found to be critical in osteoclast differentiation. The TRANCE-RANK interaction regulates the expression of the anti-apoptotic molecule Bcl-xL. Applications Reported: The MIH24 antibody has been reported for use in flow cytometric analysis, and ELISA. Applications Tested: The MIH24 antibody has been tested by flow cytometric analysis of human RANKL transfected cells. This can be used at less than or equal to 1 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- MIH24
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- 4° C
Submitted references Soluble rank ligand produced by myeloma cells causes generalised bone loss in multiple myeloma.
Expression of membrane-bound and soluble receptor activator of NF-kappaB ligand (RANKL) in human T cells.
Buckle CH, De Leenheer E, Lawson MA, Yong K, Rabin N, Perry M, Vanderkerken K, Croucher PI
PloS one 2012;7(8):e41127
PloS one 2012;7(8):e41127
Expression of membrane-bound and soluble receptor activator of NF-kappaB ligand (RANKL) in human T cells.
Kanamaru F, Iwai H, Ikeda T, Nakajima A, Ishikawa I, Azuma M
Immunology letters 2004 Jul 15;94(3):239-46
Immunology letters 2004 Jul 15;94(3):239-46
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Staining of non-transfected (left) and human RANKL transfected cells (right) with 0.25 µg of Mouse IgG2b kappa Isotype Control Purified (Product # 14-4732-82) (open histogram) or 0.5 µg of Anti-Human CD254 (RANK Ligand) Purified (filled histogram) followed by Anti-Mouse IgG FITC (Product # 11-4011-85). Total viable cells were used for analysis.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 5 RPMI-8226/hRANKL/eGFP cells express hRANKL. A. End-point PCR demonstrating that human RANKL mRNA is present in RPMI-8226/hRANKL/eGFP cells (labeled hRANKL). hRANKL mRNA was not detected in the corresponding eGFP-only control cell line (labeled eGFP), or in wild-type cells (labeled WT). No product was amplified in the absence of RNA (labelled no RNA) or the superscript enzyme (labeled SS-). B. Western Blot analysis detected the hRANKL monomer (~34 kDa) in culture supernatant from RPMI-8226/hRANKL/eGFP cells - red outline. (i) Wild-type and eGFP-only expressing cell controls are included, and the rhRANKL antibody-control is also shown (~23 kDa). (ii) Ribosomal RNA loading controls were included. C. Flow cytometric analysis of the RPMI-8226/hRANKL/eGFP-expressing cells. Representative plots showing cells stained with anti-hRANKL-PE antibody (eBioScience, 1/50 dilution) (transparent profile) and isotype control (solid profile).