Cat. No.
MABL-1322
Application
immunogold labeling, IP, ELISA, IF
Isotype
Engineer antibody
Species Reactivity
Pigeon, Rat, Mouse
Clone No.
2.7D5
From
Recombinant Antibody
Specificity
This antibody is specific for pigeon cytochrome c (CYT) and cross-reacts with mouse CYT. Specifically, it recognizes the region around residue 62 of native mouse CYT.
Alternative Names
CYCS; CYC; HCS; THC4; Cytochrome c, somatic; Cytochrome c; Cyt c
UniProt
P62897
Immunogen
The original antibody — a secondary immune response antibody — was generated by immunizing BALB/c mice with pigeon cytochrome C covalently coupled to ovalbumin using glutaraldehyde (CYT-OVA) and later challenging them with the same immunogen (pigeon CYT-OVA).
Application Notes
The binding specificity of the original format of this antibody (mouse IgG2b) was evaluated by indirect ELISA, and its binding affinity was determined by competitive ELISA. It was found to bind Pigeon CYT ~5.6 times better than Mouse CYT (Liu et al., 2000; PMID: 11257306). This antibody was used in the detection of intracellular mouse cytochrome c (cyt c) in live DO11.10 T hybridoma cells by immunofluorescent (IF) labeling. Specifically, it was found to recognize the surface around residue 62 of cyt c and bound to cyt c in fixed and permeabilized cells. Furthermore, it was used in indirect IF staining of cyt c in post-apoptotic cells and immunoprecipitation (IP) of cyt c extracted from live, apoptotic, and necrotic DO11.10 cells (Jemmerson et al., 1999; PMID: 10090746). This antibody was used in immunogold labeling to determine the subcellular localization of cytochrome-c in rat pancreatic acinar cells (Soltys et al., 2001; PMID： 11319839).
Antibody First Published
Liu et al. Kinetic and genetic bases for the heteroclitic recognition of mouse cytochrome c by mouse anti-pigeon cytochrome c monoclonal antibodies Mol Immunol. 2000 Oct;37(14)：847-59. doi： 10.1016/s0161-5890(01)00003-7 PMID：11257306
Note on publication
The original publication investigated the kinetic and genetic factors underlying the recognition of mouse cytochrome c by mouse anti-pigeon cytochrome c monoclonal antibodies, exploring the heteroclitic response during the immune response and identifying somatic mutations and Ig gene joints that affected the binding affinity and rates of the antibodies towards both mouse and pigeon cytochrome c.
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.
