Anti-Muscarinic Receptor M2, amino acids 225-359 (Cholinergic Muscarinic Receptor M2, Chrm2), clone

Applications:, Suitable for use in Immunohistochemistry and Immunocytochemistry. Does not work on paraffin embedded tissues, and Immunoblotting. Other applications have not been tested. Recommended Dilution: Immunohistochemistry (4% paraformaldehyde fixed tissue): 1-5ug/ml, Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. , , IMMUNOHISTOCHEMISTRY PROTOCOL, Use on 30 um, free floating, 4% paraformaldehyde fixed rat brain tissue. All steps are performed under constant agitation. , 1. 3 x 10 minute washes in TBS (with or without 0.25% Triton). , 2. Incubate for 30 minutes in TBS with 3% serum (same as host from secondary antibody). , 3. Incubate M9520-13B diluted appropriately in TBS with 1% serum (same as host from secondary antibody) (with or without 0.25% Triton) for 2 hours at RT followed by 16 hours at 4°C. , 4. 3 x 10 minute washes in TBS. , 5. Incubate with secondary antibody diluted appropriately in TBS with 1% serum (same as host from secondary antibody). , 6. 3 x 10 minute washes in TBS. , 7. ABC Elite (1:200) in TBS. , 8. 2 x 10 minute washes in TBS. , 9. 1 x 10 minute wash in phosphate buffer (no saline). , 10. DAB reaction with 0.06% NiCl added for intensification. , 11. 2 x 10 minute washes in PBS. , 12. 1 x 10 minute wash in phosphate buffer (no saline).