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Product Info Summary
| SKU: | A01192-4 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Choline Acetyltransferase/CHAT Antibody Picoband®
View all Choline Acetyltransferase/ChAT Antibodies
SKU/Catalog Number
A01192-4
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Choline Acetyltransferase/CHAT Antibody Picoband® catalog # A01192-4. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-Choline Acetyltransferase/CHAT Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01192-4)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human Choline Acetyltransferase/CHAT recombinant protein (Position: T25-K731).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A01192-4 is reactive to CHAT in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
83 kDa
Calculated molecular weight
82536 MW
Background of Choline Acetyltransferase/ChAT
Choline acetyltransferase (commonly abbreviated as ChAT, but sometimes CAT) is a transferase enzyme responsible for the synthesis of the neurotransmitter acetylcholine. In humans, the choline acetyltransferase enzyme is encoded by the CHAT gene. This gene product is a characteristic feature of cholinergic neurons, and changes in these neurons may explain some of the symptoms of Alzheimer's disease. Polymorphisms in this gene have been associated with Alzheimer's disease and mild cognitive impairment. Mutations in this gene are associated with congenital myasthenic syndrome associated with episodic apnea. Multiple transcript variants encoding different isoforms have been found for this gene, and some of these variants have been shown to encode more than one isoform.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A01192-4 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human placenta whole cell, human U-87MG tissue
IHC: human placenta tissue
ICC/IF: U20S cell
FCM: SiHa cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of CHAT using anti-CHAT antibody (A01192-4).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human placenta whole cell lysates,
Lane 2: human U-87MG tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CHAT antigen affinity purified polyclonal antibody (Catalog # A01192-4) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CHAT at approximately 83KD. The expected band size for CHAT is at 83KD.
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Figure 2. IHC analysis of CHAT using anti-CHAT antibody (A01192-4).
CHAT was detected in paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CHAT Antibody (A01192-4) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
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Figure 3. IF analysis of CHAT using anti-CHAT antibody (A01192-4).
CHAT was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-CHAT Antibody (A01192-4) overnight at 4°C. DyLight®488 conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Figure 4. Flow Cytometry analysis of SiHa cells using anti-CHAT antibody (A01192-4).
Overlay histogram showing SiHa cells stained with A01192-4 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CHAT Antibody (A01192-4, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For CHAT (Source: Uniprot.org, NCBI)
Gene Name
CHAT
Full Name
Choline O-acetyltransferase
Weight
82536 MW
Superfamily
carnitine/choline acetyltransferase family
Alternative Names
Choline O-acetyltransferase (EC:2.3.1.6); CHOACTase; ChAT; Choline acetylase; CHAT CHAT CHOACTASE, CMS1A, CMS1A2, CMS6 choline O-acetyltransferase choline O-acetyltransferase|acetyl CoA:choline O-acetyltransferase|choline acetylase
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on CHAT, check out the CHAT Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for CHAT: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-Choline Acetyltransferase/CHAT Antibody Picoband® (A01192-4)
Hello CJ!
A01192-4 has been cited in 3 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Icariin Promotes Survival, Proliferation, and Differentiation of Neural Stem Cells In Vitro and in a Rat Model of Alzheimer’s Disease
Pre-Injection of Small Interfering RNA (siRNA) Promotes c-Jun Gene Silencing and Decreases the Survival Rate of Axotomy-Injured Spinal Motoneurons in Adult Mice
Wu R, Gu B, Zhao X, Tan Z, Chen L, Zhu J, Zhang M. Hum Cell. 2013 Mar;26(1):19-27. Doi: 10.1007/S13577-011-0022-3. Epub 2011 Jun 15. Derivation Of Multipotent Nestin(+)/Cd271 (-)/Stro-1 (-) Mesenchymal-Like Precursors From Human Embryonic Stem Cel...
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