概述

  • 产品名称
    Choline/Acetylcholine Assay试剂盒
  • 样品类型
    Cell culture supernatant, Urine, Serum, Plasma, Cell culture extracts, Other biological fluids, Tissue Extracts
  • 检测类型
    Quantitative
  • 灵敏度
    > 0.01 nmol/well
  • 范围
    0.01 nmol/well - 5 nmol/well
  • 检测时间
    0h 40m
  • 种属反应性
    与反应: Mammals, Other species
  • 产品概述

    Choline/Acetylcholine Assay Kit (ab65345) provides a simple and sensitive means for quantifying Choline and Acetylcholine by either a colorimetric or fluorometric method in samples such as blood, cells, culture media, fermentation media, etc.


    In the choline / acetylcholine assay protocol, free choline is oxidized to betaine, via the intermediate betaine aldehyde. The reaction generates products which react with the Choline Probe to generate color (λ= 570 nm), and fluorescence (Ex/Em 535/587 nm).


    Acetylcholine can be converted to choline by adding acetylcholinesterase to the reaction to measure total choline (choline + acetyl choline). The amount of acetyl choline is calculated by subtracting choline from total choline.


    There is no need for pretreatment or purification of samples. The kit can detect ~10 pmol-5 nmol of choline or acetylcholine.


    Cells should be cultured in choline chloride free medium.


    Choline / acetylcholine assay protocol summary:
    - add samples and standards to wells
    - add choline reaction mix with, or without, acetylcholinesterase, and incubate for 30 min at room temp
    - analyze with microplate reader

  • 说明

    If you need to purchase additional Choline Probe for this product, please see ab139505.

性能

图片

  • Acetylcholine levels from mouse brain tissue was measured using Acetylcholine assay kit (ab65345). Normal mice were anesthetized with thiopental sodium at 20minutes after the end of  electroacupuncture (EA) treatment to determine levels of ACh (Figure A). Immunohistochemical staining  mAChR (muscarinic acetylcholine receptor) M3 in mouse cerebral cortex tissue 20 minutes after the end of EA treatment (Image B).

  • Choline measured fluorometrically in mouse tissue lysates showing quantity (nmol) per mg protein of tested sample

  • Choline measured fluorometrically in cell lysates showing quantity (pmol) per 1 mln of tested cells

  • Choline measured fluorometrically in biological fluids showing quantity (nmol) per mL of tested sample

实验方案

文献

This product has been referenced in:
  • Garrido-Gil P  et al. Aging-related dysregulation in enteric dopamine and angiotensin system interactions: implications for gastrointestinal dysfunction in the elderly. Oncotarget 9:10834-10846 (2018). Functional Studies . Read more (PubMed: 29541380) »
  • van Loo B  et al. Structural and Mechanistic Analysis of the Choline Sulfatase from Sinorhizobium melliloti: A Class I Sulfatase Specific for an Alkyl Sulfate Ester. J Mol Biol 430:1004-1023 (2018). Read more (PubMed: 29458126) »
See all 10 Publications for this product

客户评价及客户问答

1-10 of 17 Abreviews or Q&A

Question
Answer

J’ai contacté le laboratoire concernant les limites de détection de ce kit et de sa sensibilité.
Comme montré sur la courbe standard pour la détection colorimétrique, la limite de détection inferieure est 1 nmol/puit.

En ce qui concerne la sensibilité de cette méthode de détection, nous pouvons déterminer la DO jusqu’à ˜0.4, donc 0.5nmol peut facilement entre mesurer avec une détection colorimétrique.

Read More

Answer

I suggest running a time course to see how much time is needed for your samples to show a linear increase and then plateau off. It is recommended to choose the final incubation time within this linear increase regime.
Also, please note it is important to measure the standards at the end point, at the same time measurements of the samples are taken.

Read More

Answer

ab65345 cannot be used for in-cell assay. It is meant for cell lysates.

Read More

Answer

Thank you for your email and your interest in our products.

I can confirm that you are right. It will be necessary to set up 2 different sets of wells for measuring the total choline and the free choline in this assay.

Therefore, to measure the acetylcholine, it will be necessary to set up duplicate wells for the total choline in the samples and duplicate wells for the free choline in the same samples. I can confirm that the acetylcholine would be the total choline - free choline.

I hope this is of help to you. Please do not hesitate to contact me again if you need any further information.

Read More

Answer

Thank you for contacting us.

We have not tested this kit with rat samples. However this kit should work as it does not contain any human specific antibody. The species is irrelevant for this product.

It will also be a good idea to include human samples as positive control.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Answer

Thank you for contacting us.

All of the information and images we have for each of our products is listed on the online datasheet for your convenience. These are updated as soon as any new information is brought to our attention. Unfortunately we don’t have any references for this kit in Rat synaptosomes.

I am very sorry I couldn’t help you further. Please do not hesitate to contact us if you need any other advice or information.

Read More

Answer

Thank you very much for contacting Abcam. Your details has been forwarded to me by my colleague Peter.

The kit ab65345 have a detection range of 0.01 nmol/well to 5 nmol/ml; expression outside of this range will be undetectablewith the kit.

I am sure the kit will provide best results with suitable samples. However if the kits fails please let us know wewill behappy to help.

Best of luck with your research.

Read More

Answer

Thank you for pointing this out. The datasheet is somewhat mis-leading with regards to the 500-fold range that is specified, and also in error with regards to the notation. I apologize for the confusion.

The specified range uses the low value for the fluorometric assay and the high value for the colorimetric assay. A 500-fold range of detection can be achieved but not with a single reading.

The values of the range should specified as amounts per well, that is, 0.01 nmol - 5 nmol, not as molar concentrations (nM). Thank you again for spotting this; we will fix it.

Please let me know if you have any other questions or concerns.

Read More

Question
Answer

Tissue culture SN can be used with this assay. Add up to 50 µl of the SN to wells and make up the final volume in all wells to 50 µl with the assay buffer. Continue with the protocol.

Read More

Answer

Thank you for your email. I am sorry to hear that you have been experiencing problems with this kit.

I have checked the details you have kindly provided; however I would like to ask few questions in order to understand the problem;

- Could you provide the order number of the kit?
- The kit worked first time and then reading inverted. How long the kit reagents were stored?
- Have you checked carefully if the dilutions are not inverted?
- Could you send us raw data?
- Could you specify the protocol steps followed?

The readings are inverted for both standard and sample so it may well be that error may have happened in dilution of the standard and sample so your answers will be very helpful to know this anomaly.

I will look forward to hear from you soon soon.

Read More

1-10 of 17 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

注册