I am happy to share these 2 reagents on a collaborative basis following simple MTA execution. These rabbit monoclonals are much more sensitive than mouse clone 9E10. They work beautifully for formalin/paraffin IHC, ideal for detecting transgenes in mice.
Rabbit monoclonal anti-myc tag IgG
Antigen: EQKLISEEDL peptide
Concentrated supernatant: 10-15µg IgG/300µl
Contains 50% glycerol
Store at -20°C
Western Blotting: 1:2000-1:20000
Immunohistochemistry, formalin/paraformaldehyde fixed, paraffin sections:
Tissues were processed with an automated processor and 5µm sections were mounted on slides and baked at 60C for one hour prior to deparaffinization.
Antigen retrieval (25 min in vegetable steamer, citrate buffer, pH 6.0) was performed followed by blocking with 2% normal goat serum and 1% bovine serum albumin (BSA) in PBS, followed by incubation with the avidin/biotin block for 15 min (Sp2001, Vector Laboratories, Burlingame, CA). Slides were washed four times for 10 minutes between incubations with PBS containing 0.5% Tween-20 (PBS-T). Antibody was diluted in 2% normal goat serum and 1% BSA in PBS and incubated at 4°C overnight. Biotin conjugated anti-rabbit IgG (111-065-144, Jackson Immunologicals, West Grove, PA) was used. Endogenous peroxidase was blocked by incubation with 3% hydrogen peroxide in PBS for 15 min. The ABC Elite kit (Pk6100, Vector Laboratories, Burlingame, CA) was used with diaminobenzidine as substrate. Counterstaining was carried out with Gill1 Hematoxylin (72404, Richard Allen, VWR) for 5-10 seconds prior to dehydration and mounting with Permount (Fisher) under coverslips.
Clone ID: Vli-47
Host species: rabbit
Immunoglobulin isotye: IgG
Species Cross-reactivity: n/a
Immunohistochemistry (formalin fixed, paraffin embedded, no antigen retrieval required)
Molecular Wt: n/a
Specificity: A synthetic peptide corresponding to the myc epitope tag (EQKLISEEDL) was used as immunogen. The antibody recognizes C terminal myc epitope tags only. It does not recognize N terminal myc epitopes.
IHC: 1:1000-10000 (paraffin-embedded, formaldehyde fixed)