CHAT Polyclonal antibody

CHAT Polyclonal Antibody for WB, IHC, IF-P, IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF-P, IP, ELISA

Conjugate

Unconjugated

Publications

4

Cat no : 24418-1-AP

Synonyms

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

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Validation Data Gallery

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  WB analysis of rat brain using 24418-1-AP

  rat brain tissue were subjected to SDS PAGE followed by western blot with 24418-1-AP (CHAT Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of human placenta using 24418-1-AP

  human placenta tissue were subjected to SDS PAGE followed by western blot with 24418-1-AP (CHAT Antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse brain using 24418-1-AP

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 24418-1-AP (CHAT Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  IP experiment of rat brain using 24418-1-AP

  IP result of anti-CHAT (IP:24418-1-AP, 4ug; Detection:24418-1-AP 1:2000) with rat brain tissue lysate 1880 ug.

• 

  IHC staining of rat brain using 24418-1-AP

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 24418-1-AP (CHAT antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat brain using 24418-1-AP

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 24418-1-AP (CHAT antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse brain using 24418-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 24418-1-AP (CHAT Antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse brain using 24418-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 24418-1-AP (CHAT Antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat brain using 24418-1-AP

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 24418-1-AP (CHAT antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of mouse brain using 24418-1-AP

  Immunofluorescent analysis of (4% PFA) fixed mouse brain tissue using 24418-1-AP (CHAT antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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Tested Applications

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

Recommended dilution

• Western Blot (WB): WB : 1:500-1:2000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)-P: IF-P : 1:50-1:500

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

• IF: See 4 publications below

Product Information

24418-1-AP targets CHAT in WB, IHC, IF-P, IP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: mouse
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: CHAT fusion protein Ag19550
• Full Name: choline acetyltransferase
• Calculated Molecular Weight: 748 aa, 83 kDa
• Observed Molecular Weight: 60-66 kDa, 45 kDa
• GenBank Accession Number: BC130617
• Gene Symbol: CHAT
• Gene ID (NCBI): 1103
• RRID: AB_2879536
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• 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 to be present exclusively in the cytoplasm of cholinergic neurons and anti-CHAT antibody has been considered as the best marker for cholinergic neurons. It has been observed that CHAT gene generates multiple mRNA splice variants, including peripheral type CHAT (pCHAT) and common type CHAT (cCHAT), encoding 40-55 kDa and 66-70 kDa forms of proteins, respectively.

Publications

Species	Application	Title
mouse	IF	Nat Neurosci; Aerobic glycolysis is the predominant means of glucose metabolism in neuronal somata, which protects against oxidative damage; Authors - Yao Wei
mouse	IF	bioRxiv; Expression of the ACE2 virus entry protein in the nervus terminalis reveals the potential for an alternative route to brain infection in COVID-19.; Authors - Katarzyna Bilinska
mouse	IF	Neurobiol Dis; NRF2 activation suppresses motor neuron ferroptosis induced by the SOD1G93A mutation and exerts neuroprotection in amyotrophic lateral sclerosis; Authors - Biying Yang
mouse	IF	Int Immunopharmacol; New strategy to treat spinal cord injury: Nafamostat mesilate suppressed NLRP3-mediated pyroptosis during acute phase; Authors - Yongfu Lou