Cat. No.
MABL-2579
Application
IP, neutralizing, WB, IHC
Isotype
Engineer antibody
Species Reactivity
Rat, Human, Mouse
Clone No.
Y13-259
From
Recombinant Antibody
Specificity
The antibody binds to an epitope common to all p21ras isoforms (Glu63–Arg73), a region known as loop 4. The antibody broadly reacts with ras proteins, including M-Ras and N-Ras.
Alternative Names
p21 ras protein; Ras; p21ras
UniProt
P78460; O70564
Immunogen
The original antibody was generated by immunizing rats with Ha-MuSV-transformed NRK cells. Screening of sera of individual animals was carried out to identify those with the strongest immune response against p21. Spleens were removed from the immunised animals and cell fusion of immune lymphocytes with rat myeloma cell line 210RCY3 Ag 1.2.3 (Y3) was carried out. Supernatants were harvested and assayed for antibodies to p21 by GDP binding assay or immunofluorescence on Ha-SV- transformed MDCK cells, followed by testing for the ability of the antibodies to immunoprecipitate p21.
Application Notes
The antibody was used in studies of p21ras signaling. The antibody, which recognizes ras p21 proteins specifically blocked the serum-induced mitogenic response of tissue culture cells. When the antibody was coinjected with purified ras protein into NIH 3T3 cells, it neutralized the coinjected ras protein (Mulcahy et al., 1985; PMID: 3918269). HASM, T24, SW480 and SK-N-SH cells were lysed and used for immunoprecipitation. In this procedure the protein was immunoprecipitated using this antibody. When HASM cells were microinjected with the antibody, it significantly inhibited EGF- and thrombin-induced cell proliferation (Ammit at al., 1999). The antibody detected the p21 protein by western blot analysis. Further, peptide competition was carried out to define the binding site of the antibody (Sigal et al., 1986; PMID: 2425352). This antibody was used for immunohistochemistry on human breast tissues (Agnantis et al.1986).
Antibody First Published
Furth et al. Monoclonal antibodies to the p21 products of the transforming gene of Harvey murine sarcoma virus and of the cellular ras gene family. J Virol. 1982 Jul;43(1):294-304. PMID:6287003
Note on publication
Describes the generation of monoclonal antibodies to p21 encoded by Ha-MuSV.
Size
100 μg Purified antibody.
Concentration
1 mg/ml.
Purification
Protein A affinity purified
Buffer
PBS with 0.02% Proclin 300.
Storage Recommendation
Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.
