Tested Applications
| Positive WB detected in | mouse brain tissue, mouse placenta tissue, SH-SY5Y cells, rat brain tissue, fetal human brain tissue |
| Positive IP detected in | rat brain tissue |
| Positive IHC detected in | mouse brain tissue, human brain tissue, human lung cancer tissue, rat brain tissue Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0 |
| Positive IF-P detected in | mouse brain tissue |
| Positive IF/ICC detected in | hiPSC-derived reporter spinal motor neurons, Neuro-2a cells |
Recommended dilution
| Application | 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 |
| Immunohistochemistry (IHC) | IHC : 1:200-1:800 |
| Immunofluorescence (IF)-P | IF-P : 1:50-1:500 |
| Immunofluorescence (IF)/ICC | IF/ICC : 1:250-1:1000 |
| 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. | |
Product Information
20747-1-AP targets CHAT in WB, IHC, IF/ICC, IF-P, 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 Predict reactive species |
| 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 -20oC 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.
Protocols
| Product Specific Protocols | |
|---|---|
| IF protocol for CHAT antibody 20747-1-AP | Download protocol |
| IHC protocol for CHAT antibody 20747-1-AP | Download protocol |
| IP protocol for CHAT antibody 20747-1-AP | Download protocol |
| WB protocol for CHAT antibody 20747-1-AP | Download protocol |
| Standard Protocols | |
|---|---|
| Click here to view our Standard Protocols |
Publications
What published studies show
The CHAT Polyclonal antibody (Cat# 20747-1-AP) has 81 citations. Publications appear in high-impact journals including Advanced Materials, Nature Chemical Biology, Cell Stem Cell, Redox Biology, Oncogene, Biomaterials, and Cell Reports Medicine. Research fields span neurodegeneration (Alzheimer's, ALS, Parkinson's, SMA), cholinergic signaling, spinal cord injury, neuroinflammation, gut-brain axis, cardiac autonomic remodeling, cognitive impairment, stem cell differentiation, and ferroptosis. Applications include IF, WB, and IHC across mouse, rat, and human samples.
| Species | Application | Title |
|---|---|---|
Adv Mater Directed Neural Stem Cells Differentiation via Signal Communication with Ni-Zn Micromotors | ||
Adv Mater Noninvasive Optogenetics Realized by iPSC-Derived Tentacled Carrier in Alzheimer's Disease Treatment | ||
Bioact Mater Construction of functional neural network tissue combining CBD-NT3-modified linear-ordered collagen scaffold and TrkC-modified iPSC-derived neural stem cells for spinal cord injury repair | ||
Cell Stem Cell Deriving Schwann cells from hPSCs enables disease modeling and drug discovery for diabetic peripheral neuropathy | ||
Redox Biol Choline alleviates cognitive impairment in sleep-deprived young mice via reducing neuroinflammation and altering phospholipidomic profile | ||
Nat Chem Biol Small molecule agonist of mitochondrial fusion repairs mitochondrial dysfunction |
Reviews
What customers say
Most reviewers rate this CHAT antibody highly for Western blot, with one user confirming clear staining at ~70 kDa in mouse cortex lysate at 1:2000 dilution and another reporting excellent WB and IHC results at 1:1000. One user also praised its performance in human EC cells. However, one reviewer reported unsuccessful IHC/IF staining in mouse spinal cord at 1:100 dilution despite following a standard protocol with citrate antigen retrieval. Overall, the antibody performs well in WB, while IHC/IF results may require optimization.
The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.
Sai Sindhura (Verified Customer) (08-19-2026) | CHAT antibody gives very good staining
|
Ashley (Verified Customer) (12-16-2025) | Thank you!
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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.
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Xin (Verified Customer) (01-23-2022) | Good performance in WB (around 70 kd).
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