CHAT Polyclonal antibody

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  WB analysis using 20747-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 20747-1-AP (CHAT antibody) at dilution of 1:800 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse placenta using 20747-1-AP

  mouse placenta tissue were subjected to SDS PAGE followed by western blot with 20747-1-AP (CHAT antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  IP experiment of rat brain using 20747-1-AP

  IP result of anti-CHAT (IP:20747-1-AP, 4ug; Detection:20747-1-AP 1:500) with rat brain tissue lysate 4000ug.

CHAT Polyclonal Antibody for WB, IP, ELISA

Cat No. 20747-1-AP

Publications(52)

Reviews (2)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF, IP, ELISA

Synonyms

CHOACTase, Choline acetylase, choline acetyltransferase, Choline O-acetyltransferase, EC:2.3.1.6

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Tested Applications

• Positive WB detected in: mouse brain tissue, mouse placenta tissue, rat brain tissue
• Positive IP detected in: rat brain tissue

Recommended dilution

• Western Blot (WB): WB : 1:500-1:1000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 25 publications below
• IHC: See 6 publications below
• IF: See 27 publications below

Product Information

20747-1-AP targets CHAT in WB, IHC, IF, IP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: choline acetyltransferase
• Calculated Molecular Weight: 83 kDa
• Observed Molecular Weight: 47-50 kDa, 66-70 kDa
• GenBank Accession Number: NM_020549
• Gene Symbol: CHAT
• Gene ID (NCBI): 1103
• RRID: AB_10898169
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: P28329
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

CHAT (choline acetyltransferase) belongs to the carnitine/choline acetyltransferase family. It catalyzes the reversible synthesis of acetylcholine (ACh) from acetyl CoA and choline at cholinergic synapses. Defects in CHAT are the cause of congenital myasthenic syndrome with episodic apnea (CMSEA). CHAT is known to be present exclusively in the cytoplasm of cholinergic neurons and anti-CHAT antibody has been considered as the best marker for cholinergic neurons. Human CHAT gene generates multiple mRNA splice variants (PMID: 10861222). All these variants possess start codons for translation of 70 kDa cChAT protein (isoform 1). The M variant has an additional start codon for 83 kDa cChAT protein (isoform 2), which has been found in nuclei of neuronal cells in human CNS; and the S variant has another codon for 74 kDa cChAT protein (isoform 3). Besides, CHAT includes a short exon-skipping splice variant (peripheral type CHAT) pCHAT encoding 40-55 kDa (PMID: 31040770, 29759072). This antibody can recognize both pCHAT and cCHAT.

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
mouse	IF	Adv Mater; Noninvasive Optogenetics Realized by iPSC-Derived Tentacled Carrier in Alzheimer's Disease Treatment; Authors - Yuewen Zhai
rat	IF	Cell Stem Cell; Deriving Schwann cells from hPSCs enables disease modeling and drug discovery for diabetic peripheral neuropathy; Authors - Homa Majd
human	IF	Nat Chem Biol; Small molecule agonist of mitochondrial fusion repairs mitochondrial dysfunction; Authors - Yingjie Guo
human	IF	PLoS Biol; LSM12-EPAC1 defines a neuroprotective pathway that sustains the nucleocytoplasmic RAN gradient.; Authors - Jongbo Lee
mouse	IF	Stem Cells Transl Med; Premigratory neural crest stem cells generate enteric neurons populating the mouse colon and regulating peristalsis in tissue-engineered intestine.; Authors - Huipu Yuan
mouse	IF	J Neurosci; Myocardial Infarction Causes Transient Cholinergic Transdifferentiation of Cardiac Sympathetic Nerves via gp130.; Authors - Antoinette Olivas

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Jerome (Verified Customer) (07-20-2022)

we followed this protocol : antigen retrieval buffer: citrate pH 6 (DAKO S1699) wash in TBS 1X Blocking buffer TBS1x +Serum Goat 5%, 1H at RT Primary and secondary antibody diluted in TBS1X Wash primary antibody TBS1X 0.2%tween Secondary antibody GAR 594 ref A11012 Wash secondary antibody TBS1X 0.2%tween Unfortunately, it doesn't work under these conditions.

• Applications: Immunohistochemistry, Immunofluorescence
• Primary Antibody Dilution: 1/100
• Cell Tissue Type: Mouse spinal cord

FH

Xin (Verified Customer) (01-23-2022)

Good performance in WB (around 70 kd).

• Applications: Western Blot
• Primary Antibody Dilution: 1:2000
• Cell Tissue Type: mouse cortex lysate

CHAT Antibody Western Blot validation (1:2000 dilution) in mouse cortex lysate (Cat no:20747-1-AP)