Application

Immunohistochemistry: 1:1,000-1:2,000

Immunoblotting

Optimal working dilutions must be determined by end user.

IMMUNOHISTOCHEMISTRY PROTOCOL FOR AB1582

STAINING OF GUINEA PIG ILEUM

1) Pin the ileum out flat and tight.

2) Fix overnight at 4°C in modified Zamboni′s (2% paraformaldehyde with 15% saturated picric acid in phosphate buffered saline).

3) Clear in DMSO, 100%, 3 times for 10 minutes each, rinse in PBS.

4) Dissect off ALL circular muscle.

5) Preincubate with 10% normal donkey serum.

6) Incubate in anti-ChAT (AB1582) diluted 1:1,000-1,2000 with 10% normal donkey serum over night at 4°C on a shaker.

7) Incubate with Donkey anti-Sheep IgG-Cy3 labeled secondary antibody (Catalog Number AP184C) at a dilution of 1:200 for 2 hours.

IMPORTANT NOTE: It is essential to remove ALL circular muscle.

Research CategoryNeuroscience

This Anti-Choline Acetyltransferase Antibody is validated for use in IH, WB for the detection of Choline Acetyltransferase.

Research Sub CategoryNeurotransmitters & ReceptorsNeuronal & Glial Markers

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Immunogen

A 22 amino acid peptide from porcine ChAT, coupled to KLH. The sequence is H-GLFSSYRLPGHTQDTLVAQKSS-NH2.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Physical form

Sheep serum. Lyophilized, no preservatives. Reconstitute with 100 µL of sterile distilled water. Centrifuge to remove any residue.

Specificity

Choline Acetyltransferase (ChAT). Will stain cholinergic neurons in rat, guinea-pig and rabbit central and peripheral nervous systems.

Storage and Stability

Maintain at -20 to -70°C in undiluted aliquots for up to 12 months. Avoid repeated freeze/thaw cycles. Glycerol (1:1) can be added for additional stability.