Depositors Institution: Johns Hopkins School of Medicine
Positive Tested Species Reactivity: Avian, Canine, Fish, Human, Mouse, Porcine, Rat, Torpedo, Zebrafish
Depositors Notes: Recognizes a cytosolic epitope on the alpha-subunit of the Na, K-ATPase, alpha-1 isoform. The epitope appears to be linear, and located on the N-terminal region between aa 27-55. Weak or no binding to sheep and Xenopus. Works great for Immunoblotting. Immunochemistry: good on cryosections; poor for lightly-fixed cells in tissue culture; seems to require denaturation of Na, K-ATPase. This hybridoma was made together with alpha-7C, described in [PMID 2831227].
Additional Information: Broad species specificity [PMID 8798546]. This antibody doesn't appear to work on conventionally fixed tissue, and stains fixed sections of the rat kidney faintly, but this staining is considerably increased by prior treatment of the sections with SDS [PMID 8798546]. RRID: AB_528092
Recommended Applications: ELISA, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, 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:
a6F was deposited to the DSHB by Fambrough, D.M. (DSHB Hybridoma Product a6F)
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.