United Kingdom (UK)
Recombinant monoclonal antibody to AIF1. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma GT1-mAb1.
UniProt Accession Number of Target Protein: P55008
Alternative Name(s) of Target: AIF1; IBA1; allograft inflammatory factor 1; AIF-1; IRT-1; interferon gamma responsive transcript; ionized calcium-binding adapter molecule 1; Daintain; DADB-70P7.8; allograft inflammatory factor-1 splice variant Hara-1; IRT1; protein G1
Immunogen: This antibody was obtained by sequencing a polyclonal goat IgG raised against a peptide with the sequence C-TGPPAKKAISELP.
Specificity: This antibody is specific to EBP05419, an immunizing peptide with the sequence C-TGPPAKKAISELP. This peptide was used in the production of the polyclonal Goat Anti-AIF1 antibody from which this antibody is derived.
Application Notes: This antibody (GT1-mAb1) was one of two monoclonal antibodies discovered by analyzing a polyclonal antibody (GT-1) sample. Briefly, de novo sequencing of GT-1 resulted in the full-length sequences of two distinct monoclonal antibodies (GT1-mAb1 and GT1-mAb2) in both framework and CDR regions. Both antibodies were recombinantly expressed by Absolute Antibody in HEK293 cells and tested in various assays. GT1-mAb1 was tested against the peptide antigen in an ELISA, and it showed activity comparable to that of the original polyclonal antibody. GT1-mAb1 was found to bind the target peptide in a non-competitive manner with GT1-mAb2 while being efficiently blocked by the original polyclonal antibody, proving that both monoclonal antibodies bind distinct epitopes and that GT1-mAb1 possess binding characteristics of the input polyclonal antibody. Furthermore, GT1-mAb1 was compatible with immunofluorescence (IF), flow cytometry (FC), and Western blot (WB), with an FC signal observedly more distinct than that of GT1-mAb2.
Antibody first published in: PMID:
Immunofluorescence staining of U937 cells with anti-AIF1 antibody GT1-mAb1 (Ab03986 (previously cAb7701)). Immunofluorescence analysis of paraformaldehyde fixed U937 cells on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of GT1-mAb1 (Ab03986-24.1) (10ug/ml) for 1h followed by Alexa Fluor® 488 secondary antibody (2ug/ml), showing membrane staining. The nuclear stain is DAPI (blue). Panels show, from left-right, top-bottom, Ab03986-24.1, DAPI, merged channels and an isotype control. The isotype control was an anti-fluorescein antibody (Ab00102-24.1) followed by staining with Alexa Fluor® 488 secondary antibody.
Western blot using anti-AIF1 antibody GT1-mAb1 (Ab03986 (previously cAb7701)). RAW264.7 (A) (2µg/ml), U937 (B) (2µg/ml) cells lysate, and mouse brain (C) (2µg/ml) tissue lysate (35µg protein in RIPA buffer) were resolved on an SDS-PAGE gel, and blots were probed with the chimeric rabbit version of GT1-mAb1 (Ab03986-24.1) at 2µg/ml before detection using an anti-rabbit secondary antibody. A primary incubation of 1 hour was used, and protein was detected by chemiluminescence.
Flow cytometry using anti-AIF1 antibody GT1-mAb1 (Ab03986). Paraformaldehyde fixed human peripheral blood monocytes permeabilized with 0.5% Triton were stained with the isotype control antibody (Ab00102-24.1; black line) or the rabbit IgG version of GT1-mAb1 (Ab03986-24.1, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000, and the cells were analyzed using a FACSCanto flow-cytometer.