Depositors Institution: Michigan State University, Dept. of Biochemistry & Molecular Biology
Positive Tested Species Reactivity: Rat
Depositors Notes: Monoclonal 2A4 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. The epitope recognized is a conformational 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].
Additional Information: The antibody is reactive with the native enzyme (ELISA) but not the denatured form of the enzyme and thus is not useful for immunoblots. The functional effects of this antibody inhibits rat Type I hexokinase activity to >95%, the effect being on V max with no significant effect of Kms for either glucose or ATP [Smith and Wilson (1992) Arch. Biochem. Biophys. 292: 165-178. RRID:AB_528280
Recommended Applications: ELISA, Function Blocking
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:
2A4 was deposited to the DSHB by Wilson, J.E. (DSHB Hybridoma Product 2A4)
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.