Depositors Institution: 301 Biochemistry, Dept. of Biochemistry & Molecular Biology, Michigan State University
Positive Tested Species Reactivity: Rat
Depositors Notes: The antibody is reactive with the native enzyme (ELISA) but not the denatured form of the enzyme and thus is not useful for immunoblots. Binding of the antibody does not affect the kinetic properties of the rat Type I isozyme [Smith and Wilson (1992) Arch. Biochem. Biophys. 292: 165-178]. The epitope recognized is a ÒconformationalÓ (or ÒdiscontinuousÓ) epitope, and immunoreactivity is affected by conformational changes in the Type I hexokinase, e.g., as a result of binding certain ligands [Smith and Wilson (1992) Arch. Biochem. Biophys. 292: 165-178]. rat; crossreactivity with Type I isozyme from other mammalian species has not been investigated. Monoclonal 1B2 recognizes an epitope in the C-terminal half of rat Type I hexokinase. Crossreactivity with the Type II or Type III isozymes is thought to be unlikely. Crossreactivity with the Type II or Type III isozymes is thought to be unlikely.
Additional Information: The origin of the antibody is purified from rat brain. You can find details of the immunization and subsequent fusion protocol are found in Finney et al. (1984) J. Biol. Chem. 259: 8232-8237. Immunization of 100 micrograms of purified rat type I hexokinase was injected intraperitoneally and the adjuvant is freunds. RRID:AB_528279
Recommended Applications: ELISA, Western Blot
These hybridomas were created by your colleagues. Please acknowledge the hybridoma contributor and the Developmental Studies Hybridoma Bank (DSHB) in the Materials and Methods of your publications. Please email the citation to us.
For your Materials & Methods section:
1B2 JW was deposited to the DSHB by Wilson, J.E. (DSHB Hybridoma Product 1B2 JW)
Storage and Handling Recommendations
Although many cell products are maintained at 4°C for years without loss of activity, shelf-life at 4°C is highly variable.
To ensure retention of antibody activity, we recommend aliquotting the product into two parts:
1) a volume of antibody stored at 4°C to be used within two weeks.
2) the remaining product diluted with an equal volume of molecular grade glycerol and stored at -20°C.
While optimal Ig concentration for an application will vary, a good starting concentration for immunohistochemistry (IHC),
immunofluorescence(IF) and staining is 2-5 µg/ml.
For Western blots, the concentration is decreased by one order of magnitude (that is, 0.2-0.5 µg/ml).
All cell products contain the antimicrobial ProClin. See our FAQ section for additional information.