United Kingdom (UK)
Recombinant monoclonal antibody to Ly-6C (T cell activating). Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma HK1.4.
UniProt Accession Number of Target Protein: P0CW02
Alternative Name(s) of Target: Lymphocyte antigen 6C1; Ly6c1
Immunogen: The antibody was raised by immunizing a Lewis rat by a series of four monthly i.p. injections of approximately 30 X 10" L3 cloned CTL cells.
Specificity: This antibody is specific for Ly6 and shows no cross reactivity. Ly6 shows acetylcholine receptor binding activity.
Application Notes: A flow cytometry experiment on mouse cells from L-gD-1, an L cell line that expresses HSV type 1 gD, and L-gI-1, an L cell line that expresses HSV type 1 gI was preformed. In this experiment this antibody was used for reverse blocking (Johnson et al, 1993; pmid:8104217). Th1 clones from either wild-type or fyn-/- mice were used in an experiment where it was determined if this antibody could induce lysis (Lancki et al, 1995; pmid:7493751). Flow cytometry was preformed on cells from C57BL/6, DBAI2, NJ. BALB/c, PIJ, PL/J. AKRIJ. RIII, SM/J, SJL, and CBA mice using this antibody (Havran et al, 1988; pmid:3257767). Immunofluorescence was preformed on rat EL4J cells using this antibody (Fleming et al, 1993; pmid:8360469). EL4J and EUJ-Ly-6G cell lines were immunoprecipitated with this antibody (Fleming et al, 1993; pmid:8360469). Neonatal murine calvaria-derived osteoblasts were stained for flow cytometry using this antibody (Horowitz et al, 1994; pmid:7520861).
Antibody first published in:
Havran et al. Characterization of an anti-Ly-6 monoclonal antibody which defines and activates cytolytic T lymphocytes J Immunol. 1988 Feb 15;140(4):1034-42. PMID:3257767
Note on publication:
The paper describes the generation of the antibody is rats by immunizing them with L3 cloned CTL cells.
Immunofluorescence staining of mouse splenocytes with anti-Ly-6C (T cell activating) antibody HK1.4 (Ab02560). Immunofluorescence analysis of paraformaldehyde-fixed mouse splenocytes on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of HK1.4 (Ab02560-23.0) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing membrane staining in a subset of cells. The nuclear stain is DAPI (blue). Panels show, from left-right, top-bottom, Ab02560-23.0, DAPI, merged channels, and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.