Depositors Notes: To expose the core protein epitope, the tissue must be reduced and alkylated. Treat the tissue with 10mM dithiothreitol in TBS pH 7.4 for 2 hr at 37ºC for 2 hr. Follow by alkalation with 40 mM iodoacetamide in PBS for 2 hr.
Immunogen: reduced and alkylated rat chondrosarcoma proteoglycan monomer
Additional Information: The amount of antigent used in immunization was injected at six sites with 0.5 ml of protein (200-800 ?g protein/ml) and 0.5 ml complete Freund’s adjuvant on day 1; 0.5 ml Freund’s incomplete adjuvant on day 3; 0.5 ml Dulbecco’s PBS on days 6, 9, and 12; on day 14. The mouse was sacrificed for the route of immunization.
Positive Tested Species Reactivity: Bovine, Canine, Chicken, Fish, Human, Pig, Rat
Recommended Applications: Immunohistochemistry
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:
12/21/1-C-6 was deposited to the DSHB by Caterson, B. (DSHB Hybridoma Product 12/21/1-C-6)
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.