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  1. Link

    alanine-transaminase-activity-assay-kit-colorimetricfluorometric-ab105134.pdf

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Signal Transduction Metabolism Amino Acids
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Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (2)Q&A (17)References (67)

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Determination of ALT activity using ab105134 Alanine Transaminase Activity Assay Kit
  • Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)
  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)
  • Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)
  • Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)
  • Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)
  • Functional Assay - ab105134
  • Functional Studies - Alanine Transaminase Activity Assay Kit (105134)
  • Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)
  • Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)

Key features and details

  • Assay type: Enzyme activity (quantitative)
  • Detection method: Colorimetric/Fluorometric
  • Platform: Microplate reader
  • Assay time: 1 hr 20 min
  • Sample type: Cell culture media, Cell culture supernatant, Other biological fluids, Plasma, Serum, Tissue Extracts, Urine
  • Sensitivity: 10 mU/well

こちらの製品もご検討ください

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Aspartate Aminotransferase Activity Assay Kit (ab105135)
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ELISA
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Human ALT ELISA Kit (ab234578)

関連製品

製品の概要

  • 製品名

    Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric)
    Alanine Transaminase キット 製品一覧
  • 検出方法

    Colorimetric/Fluorometric
  • サンプルの種類

    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell culture media
  • アッセイタイプ

    Enzyme activity (quantitative)
  • 検出感度

    > 10 mU/well
  • 全工程の試験時間

    1h 20m
  • 製品の概要

    Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) ab105134 is a rapid and simple assay used to quantify alanine transaminase (ALT) activity in mammalian samples.


    In the ALT assay protocol, ALT transfers an amino group from alanine to α-ketoglutarate; producing pyruvate and glutamate. The pyruvate is detected in a reaction that converts a nearly colorless probe to a form that is colored (ODmax = 570 nm) and fluorescent (Ex/Em = 535/587 nm).


    The kit has a detection limit of 10 mU per well.


    ALT assay protocol summary:
    - add samples and standards to wells
    - add reaction mix and incubate for 10 min at 37ºC
    - analyze every 2-3 min for 60 min with microplate reader in kinetic mode at 37ºC

  • 特記事項

    This product is manufactured by BioVision, an Abcam company and was previously called K752 Alanine Aminotransferase (ALT or SGPT) Activity Colorimetric/Fluorometric Assay Kit. K752-100 is the same size as the 100 test size of ab105134.

    Alanine transaminase is also called alanine aminotransferase or serum glutamic pyruvic transaminase (ALT, ALAT, SGPT).

  • 試験プラットフォーム

    Microplate reader

製品の特性

  • 保存方法

    Store at -20°C. Please refer to protocols.
  • 内容 ラベル 100 tests 2000 tests
    ALT Assay Buffer WM 1 x 25ml 20 x 25ml
    ALT Enzyme Mix (lyophilized) Green 1 vial 20 vials
    ALT Positive Control (lyophilized) Blue 1 vial 20 vials
    ALT Substrate (lyophilized) Orange 1 vial 20 vials
    OxiRed™ (in DMSO) Red 1 x 200µl 20 x 200µl
    Pyruvate Standard (100 nmol/µl) Yellow 1 x 100µl 20 x 100µl
  • 研究分野

    • Signal Transduction
    • Metabolism
    • Amino Acids
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Kits/ Lysates/ Other
    • Kits
    • Cell Metabolism Kits
    • Amino Acid Assays
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
  • 別名

    • Alanine Aminotransferase
    • ALT

画像

  • Determination of ALT activity using ab105134 Alanine Transaminase Activity Assay Kit
    Determination of ALT activity using ab105134 Alanine Transaminase Activity Assay KitWaller-Evans H et al., PLoS One, 8(12). Fig 1d. doi: 10.1371/journal.pone.0082825 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Liver samples from high fat diet (HFD) and standard carbohydrate diet (CHD) BALB/c and C57BL6/J mice were homogenised in an ALT assay buffer for the determination of ALT activity using ab105134. A separate batch of liver extracts was prepared and incubated in a buffer containing NP40 (5%) and supernatants containing the triglycerides were separated. Triglycerides concentration was determined on the supernatant fraction using ab65336. ALT activity and triglycerides concentration were determined by measuring OD at 570nm.

  • Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)
    Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)
    Colorimetric standard curve: mean of duplicates (+/- SD) with background reads subtracted.
  • Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)
    Functional Studies - Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric) (ab105134)Qiu Guihua, et al., Frontiers in Immunology Vol 8. 2017 pg.575, Fig 8, doi:10.3389/fimmu.2017.00575

    Serum aspartate transaminase (AST) levels were measured using ab105135 and alanine transaminase (ALT) levels were measured using ab105134. Both levels were measured at various time periods post Con A injection. Mean values ± SD are shown (n = 4). ∗P < 0.05 and ∗∗P < 0.01.

  • Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)
    Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)

    Fluorometric standard curve: mean of duplicates (+/- SD) with background reads subtracted.

  • Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)
    Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)

    Alanine transaminase measured in mouse tissue lysates showing quantity (mU) per mg of tested sample.

    Protein concentration for samples varied from 4 mg/mL to 13 mg/mL. Samples were diluted 9-27 fold and measured colorimetrically.

  • Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)
    Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)

    Pyruvate measured colorimetrically in cell lysate after 20 min and 40 min incubation time showing quantity (nmol) per 1 mln of tested cells.

  • Functional Assay - ab105134
    Functional Assay - ab105134

    Measurement of alanine transaminase in HepG2 cells (10 μg) and liver lysate (15 μg).

  • Functional Studies - Alanine Transaminase Activity Assay Kit (105134)
    Functional Studies - Alanine Transaminase Activity Assay Kit (105134)

    Pyruvate measured fluorometrically in cell lysate after 20 min and 40 min incubation time showing quantity (nmol) per 1 mln of tested cells.

  • Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)
    Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)

    Pyruvate measured in biological fluids after 20 min and 40 min incubation time showing quantity (nmol) per ml of tested sample.

  • Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)
    Functional Studies - Alanine Transaminase Activity Assay Kit (ab105134)
    Pyruvate measured in mouse tissue lysates after 20 min and 40 min incubation time showing quantity (nmol) per mg of tested sample.

プロトコール

  • Protocol Booklet

Click here to view the general protocols

データシートおよび資料

  • SDS download

  • Datasheet download

    Download

参考文献 (67)

ab105134 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab105134 は 67 報の論文で使用されています。

  • Humpton TJ  et al. Differential requirements for MDM2 E3 activity during embryogenesis and in adult mice. Genes Dev 35:117-132 (2021). PubMed: 33334825
  • Dong J  et al. Hepatocyte-specific IL11 cis-signaling drives lipotoxicity and underlies the transition from NAFLD to NASH. Nat Commun 12:66 (2021). PubMed: 33397952
  • Hwang I  et al. Retinol from hepatic stellate cells via STRA6 induces lipogenesis on hepatocytes during fibrosis. Cell Biosci 11:3 (2021). PubMed: 33407858
  • Tajan M  et al. Serine synthesis pathway inhibition cooperates with dietary serine and glycine limitation for cancer therapy. Nat Commun 12:366 (2021). PubMed: 33446657
  • de la Rosa Rodriguez MA  et al. Hypoxia-inducible lipid droplet-associated induces DGAT1 and promotes lipid storage in hepatocytes. Mol Metab 47:101168 (2021). PubMed: 33465519
View all Publications for this product

レビューと Q&A

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レビューを送る 質問を送る

1-10 of 19 Abreviews or Q&A

Super easy to use

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
Consistent with other assays from different vendors. We used colorimetric in mouse plasma samples.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Dec 09 2019

Alanine Transaminase Activity Assay Kit (Colorimetric/Fluorometric)

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
perfect kit antibodies
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Jan 24 2018

Question

Precipitation in ALT Assay buffer.

Read More

Abcam community

Verified customer

Asked on Jul 09 2018

Answer

The precipitate can be re-solubilized by gentle heating with occasional swirling. Placing in hot water bath of 50 - 60 degree for 10-15 min would be more than enough. Avoid vigorous shaking as there may be foam formation.

Read More

Abcam Scientific Support

Answered on Jul 09 2018

Question

question concernant kit ab105134.

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Abcam community

Verified customer

Asked on Feb 06 2013

Answer

J’ai eu une réponse du laboratoire qui m’a confirmé que ce kit peut être utilisé sur des échantillons humain.

Malheureusement le laboratoire n’a pas de conversion entre l’activité et la concentration d’ALT. mU est équivalent à nmol/min d’ALT et le poids moléculaire d’ALT est 54.47.

Read More

Abcam Scientific Support

Answered on Feb 06 2013

Question

Hallo!
Meine Frage bezieht sich auf Ihre Produkte
Alanine Transaminase Activity Assay Kit (ab105134)
und GST Activity Assay Kit (Colorimetric) (ab65326).
Ich möchte diese Kits nutzen, um die beiden Enzyme im Überstand meiner
Zellkulturen nachzuweisen.
Auf Ihrer website wird für diese beiden Produkte angegeben, dass sie
für den "Probentyp" Zellkulutrmedium bzw. Zellkulturüberstand geeignet
sind. Leider finde ich in der jeweiligen Anleitung (protocol booklet)
keine Angaben zur Behandlung von Zellkulturüberstand.
Darum meine Frage: Ist es sicher, dass ich diese Kits zum Nachweise
einer Enzymfreistzung ins Zellkulturmedium verwenden kann?

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Abcam community

Verified customer

Asked on Jan 25 2013

Answer



Ich möchte hiermit auch bestätigen, dass beide Kits für Zellkulturüberstand getestet und auch garantiert sind.

Sie haben Recht, dass eine Probenaufbereitung nicht gesondert erwähnt wird.

Zellkulturüberstand (frei von Zellen) kann direkt eingesetzt werden. Wir empfehlen einen unverdünnte Probe und dann einige verschiedene Verdünnungen zu testen, damit man sicherstellt, dass die Werte in den Bereich der Standardkurve fallen.

Read More

Abcam Scientific Support

Answered on Jan 25 2013

Question

Dear,

Thank you for getting back to me - before we go any further to ordering, I would like a quote on the ALT positive control..
Hope to hear from you soon

Read More

Abcam community

Verified customer

Asked on Jan 10 2013

Answer

I am sorry for the delay in getting back to you. I now have some further information to share with you.

Contrary to what I mentioned in my previous email, the L-Alanine Assay Kit (ab83394) is unlikely to be of use in the experiments you are performing. This is because an alanine transaminase is not used in the conversion of the substrate. I am sorry for any confusion caused in this. The Alanine Transaminase Activity Assay Kit (ab105134) should however be applicable if you amend the protocol.

As discussed, you may need extra ALT Positive Control as the kit is not designed for the experiments you are hoping to perform but to measure ALT activity within biological samples. I would be able to add this component to the catalogue for you if you would like. This would cost £100 for 1 vial (as supplied with the whole kit).

I hope this has been of help. Please do let me know if I can be of any further assistance.

Read More

Abcam Scientific Support

Answered on Jan 10 2013

Question



I have another question regarding this kit, if the standard reaction does not include the ALT enzyme, is the positive control supplied in this kit is the ALT enzyme, if so, then we may be able to include this enzyme in the reaction in place of the biological sample and test compounds to observe inhibition????........

Hope to hear from you soon

Kind regards

Read More

Abcam community

Verified customer

Asked on Dec 20 2012

Answer

Thank you for contacting us yesterday in regards to the Alanine Transaminase Activity Assay Kit (ab105134). I have now had a chance to have a look at this product to see if it would be suitable to use in your experiment to assess the transaminase inhibition properties of your compounds.

In short, the kit should be compatible with this but would need to be modified in order to get the results that you are looking for. The kits is intended to be used to measure the inherent alanine transaminase activity of biological samples. For example, measuring the activity from liver tissue samples. This is performed by using the provided substrates for the enzyme to work on and measuring the pyruvate generated. The enzyme comes from the biological sample tested.

You would therefore need to introduce the alanine transaminase enzyme into your samples to measure the inhibition caused by your compounds. This enzyme is provided as the ALT positive control. What you would need to do is set up the reaction with this included in each of your wells and add your inhibitors. You could then compare the inhibition by observing the difference of the ALT activity in the presence and absence of inhibitor.

As the kit is not really designed for this purpose it may require some optimisation and the quantity of ALT provided is not very high as it is only intended to be used as a single positive control. You may want to consider looking at the L-Alanine Assay Kit (ab83394) instead. This kit allows for the detection of alanine but works on the same principle as ab105134. It uses ALT to convert alanine to pyruvate which is then specifically detected.

Again, this kit is not specifically desigend for the experiment you are hoping to perform and therefore the protocol would need to be modified. In this kit, the alanine standard is provided but may again be limiting in the experiment you are hoping to perform.

What I am going to do is see if either the ALT positive control for ab105134 or the alanine standard from ab83394 may be available to purchase independently of the kits. This may help you as you would be able to run more tests with each kit. Once I have this information I will get back to you.

I hope this information has been of help. If you have any further questions in the meantime, please do not hesitate to let me know.

Read More

Abcam Scientific Support

Answered on Dec 20 2012

Question

We purchased 2 kits ab105134.
Backfround is too high to obtain an accurate reading from positive control. in both kits. Second kit is worse than the first.

Read More

Abcam community

Verified customer

Asked on Nov 15 2012

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to reassure you that this kit is tested and covered by our 6 month guarantee for. In the event that a product is not functioning as cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

As discussed on the telephone, I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.

I would appreciate if you are also able to provide any data and graphs which would help us to assess the results.

Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.



A. General Information
- Catalogue number:


- Lot number:

- Purchase order number/ order date or preferably the Abcam order number:

- How was the kit stored?


B. Description of the problem
- Are the signals you obtained with control or standard satisfactory? (Yes/No);


- Are the sample readings as expected or inconsistent?


- Are the blank readings as expected?

- Other (e.g. missing components)


C. What do you think the problem is caused by? E.g. enzyme is faulty, developer is faulty or other component that you think might be faulty.



D. Sample
- Sample Type (e.g. Cell culture supernatant, Cell lysates, Tissue lysates or Serum samples)


- Sample Species (e.g. human, mouse or rat)


- Storage (at what temperature and for how long were the samples stored?)

- Sample treatment, if any



- Sometime proteins in samples interfere with readings. Have you removed proteins?


- Sample preparation


E. Protocol used:
Did you follow the protocol exactly as given on the datasheet?


F. Optimization attempts (problem solving); please describe any deviations from the protocol.



G. Have you used the same kit successfully before?


H. Do you obtain the same results every time?


I. Additional Notes; (any additional information you would like to provide)



J. Results:

Read More

Abcam Scientific Support

Answered on Nov 15 2012

Question

Dear,



I have a question about the assaykit Alanine transaminase activity kit number: ab105134



The section measurement of the protocol is a bit unclear to me. I would like to know whether after pipeting all the samples and reaction mix in the wells I should wait around 10 minutes before measuring the first absorbance A1 at T1 time point? And if the plate at this point should still be at room temperature or already at 37C.

Then If understand correctly the plate should be further incubated at 37C for 60 minutes and then the second absorbance A2 (T2) is measured.

The main question is whether the first measurement (A1)should be done at room temperature and then afterwards the well plate is incubated at 37C.

And if I understand before measuring A1 I should just wait 10 minutes.

Furthermore the protocol described to use distilled water , however I would like to know whether RO water is also fine.



Thank you very much in advance for your answer!

Read More

Abcam community

Verified customer

Asked on Oct 11 2012

Answer

Thank you for contacting us.

You are correct, the first readin should be done at RT after 10 minutes. After this you can incubate at 37 degrees away from light before taking the second reading. We have not tested this product using RO water and cannot give you any data supporting its use.

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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Abcam Scientific Support

Answered on Oct 11 2012

Question

Customer kindly contacted us as they were experiencing troubles with this product.

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Abcam community

Verified customer

Asked on Sep 20 2012

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxxxxx

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Abcam Scientific Support

Answered on Sep 20 2012

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