| Catalog # | M9520-13B |
| 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. For long-term storage and to avoid repeated freezing and thawing, add sterile 40-50% glycerol, aliquot and store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. |
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| 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). |
| CAS Number | n/a |
| Clone Type | Monoclonal |
| Isotype | IgG2a |
| Clone No | 2Q147 |
| Host | Rat |
| Source | Human |
| Concentration | ~1mg/ml |
| Form | Supplied as a liquid in PBS, pH 7.2, 0.1% sodium azide. |
| Purity | Purified immunoglobulin |
| Immunogen | I3 loop of m2 receptor fusion protein (225-359), fused to Glutathione S-transferase. |
| Specificity | M2 muscarinic acetylcholine receptor. No reactivity with the other subtypes. Species crossreactivity: Human, monkey and rat. Expected to react with most mammalian species. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
