Muscarinic Acetylcholine Receptor M2, aa225-359 (Chrm2, CHRM-2, 7TM Receptor, FLJ43243, HM2, MGC120006, MGC120007) (Biotin)

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:|Store product at 4°C if to be used immediately within two weeks. For long-term storage, aliquot to avoid repeated freezing and thawing and store at -20°C. Aliquots are stable at -20°C for 12 months after receipt. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. 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). ||Note: Applications are based on unconjugated antibody.