United Kingdom (UK)
Recombinant monoclonal antibody to DR5. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma 304.
UniProt Accession Number of Target Protein: O14763
Alternative Name(s) of Target: TRAILR2; TNFRSF10B; TRAIL receptor 2; tumor necrosis factor receptor superfamily member 10B
Immunogen: The antibody was raised in mice homozygous for both disrupted endogenous Ig heavy chain and k light chain, harboring a chromosome 14 fragment (SC20) containing a human Ig heavy chain locus and a human Igk chain transgene (KCo5). The mice were immunised against TRAIL-R1 and TRAIL-R2.
Specificity: The antibody is specific to human DR5.
Application Notes: This antibody has shown high anti-tumour activity in mice with subcutaneously transplanted colon carcinoma cells Colo205. It has tumour-specific cell-death-inducing activity alone in tissue culture so is expected to show anti-tumour activity independent of physiological conditions of the patient. The antibody has been shown to function in ELISA and RIA.
Antibody first published in: PMID:
Western Blot using anti-DR5 antibody 304 (Ab00987). HeLa(A) (0.5µg/ml) , Caco-2(B) (0.5µg/ml) and K562(C) (0.5µg/ml) cells lysate tissue lysate (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of 304 (Ab00987-23.0) before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.
Flow-cytometry using the anti-DR5 antibody 304 (Ab00987). Caco-2 cells were stained with an isotype control (Ab00102-23.0, black line) or the rabbit-chimeric version of 304 (Ab00987-23.0, blue line) at a concentration of 1 µg/ml for 30 mins at RT. After washing, bound antibody was detected using Alexa Fluor 488® conjugated goat anti-rabbit IgG and cells analysed on a FACSCanto flow-cytometer.
Immunofluorescence staining of Caco-2 cells with anti-DR5 (Ab00987) 304 Immunofluorescence analysis of paraformaldehyde fixed Caco-2 cells stained with the chimeric rabbit IgG version of 304 (Ab00987-23.0) (1:1000 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1500 dilution), showing nuclear staining. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom Ab00987-23.0, DAPI, merged channels and an isotype control. The isotype control was anti-fluorescein antibody (Ab00102-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.