Technical Data
M9520-13B
Muscarinic Acetylcholine Receptor M2, aa225-359 (Chrm2, CHRM-2, 7TM Receptor, FLJ43243, HM2, MGC120006, MGC120007)
Description:
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 4C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile 40-50% glycerol, aliquot and store at -20C. Aliquots are stable for at least 12 months at -20C. 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.

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 4C.
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).
TypeIsotypeCloneGrade
MabIgG2a2Q147Purified
SizeStorageShippingSourceHost
100ug4C (-20C Glycerol)Blue IceHumanRat
Concentration:
~1mg/ml
Immunogen:
i3 loop of m2 receptor fusion protein (225-359), fused to Glutathione S-transferase.
Purity:
Purified immunoglobulin
Form
Supplied as a liquid in PBS, pH 7.2, 0.1% sodium azide.
Specificity:
M2 muscarinic acetylcholine receptor. No reactivity with the other subtypes. Species crossreactivity: Human, monkey and rat. Expected to react with most mammalian species.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. J. Comp. Neurology (1995) 351:339356. 2. Brain Research (1997) 778:4355. 3. Neuroscience Letters (1997) 229:8184. 4. J. Comp. Neurology (1999) 410:431443