製品の概要

  • 製品名

    Malate Assay Kit
  • 検出方法

    Colorimetric
  • サンプルの種類

    Tissue Extracts, Food samples
  • アッセイタイプ

    Quantitative
  • 検出感度

    20 µM
  • 検出範囲

    20 µM - 200 µM
  • 全工程の試験時間

    1h 00m
  • 製品の概要

    Abcam's Malate Assay Kit is an easy and sensitive assay to measure the L(-) Malate level in a variety of samples. In the assay, malate is specifically oxidized to generate a product which reacts with a substrate probe to generate color (λmax = 450 nm). The assay can detect 1~10 nmol of Malate in a 50 µl sample with a detection sensitivity ~20 µM.
    Visit our FAQs page for tips and troubleshooting.

  • 特記事項

    L(-) Malate is a TCA cycle intermediate. It plays an important role in the Calvin cycle during carbon fixation in plants. In lower organisms, malate is converted to lactate during malolactic fermentation with the formation of CO2. Malate is frequently used as an additive in the food and pharmaceutical industries, so quantitating malic acid is important in manufacturing beer, wine, cheese and fruits, among others.

  • 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
    Malate assay buffer WM 1 x 20ml
    Malate Enzyme Mix (Lyophilised) Green 1 vial
    Malate Standard (10 µmol) (Lyophilized) Yellow 1 vial
    WST Substrate (Lyophilised) Red 1 vial
  • 研究分野

  • 関連性

    L(-) Malate is a TCA cycle intermediate. It plays an important role in the Calvin cycle during carbon fixation in plants. In lower organisms, malate is converted to lactate during malolactic fermentation with the formation of CO2. Malate is frequently used as an additive in the food and pharmaceutical industries, so quantitating malic acid is important in manufacturing beer, wine, cheese and fruits, among others.
  • 別名

    • Malic acid

画像

プロトコール

参考文献

This product has been referenced in:

  • Jin L  et al. The PLAG1-GDH1 Axis Promotes Anoikis Resistance and Tumor Metastasis through CamKK2-AMPK Signaling in LKB1-Deficient Lung Cancer. Mol Cell 69:87-99.e7 (2018). Read more (PubMed: 29249655) »
  • Liu J  et al. Roles of AtpI and two YidC-type proteins from alkaliphilic Bacillus pseudofirmus OF4 in ATP synthase assembly and nonfermentative growth. J Bacteriol 195:220-30 (2013). Read more (PubMed: 23123906) »
See all 2 Publications for this product

レビューと Q&A

1-7 of 7 Abreviews or Q&A

Answer

Thank you for contacting us.

The product referred in your email (ab119693) is not intended to measure Malate in your samples, but the enzymatic activity of Malate dehydrogenase.

We do have a different kit in the catalogue, ab83391, which measures Malate in various sample types. Although we have not specifically tested it with bacteria, we predict it may work with this species.

I’d like to bring your attention to an offer we are running throughout November. If you order any primary antibody you can receive a RabMab free, whilst stocks last. Simply quote “RABMAB-XBSMG” in your next primary antibody order. More information on this offer can be found from the following link:

https://www.abcam.com/index.html?pageconfig=resource&rid=15447

I hope this helps. Otherwise do not hesitate to contact me back.

Read More

Answer

Thank you for contacting us.

Yes, this assay buffer had detergents and any bubbles will interfere with the readings. Please take care when pipeting to avoid bubbles. Should you have many bubbles please try to remove these from the well.



The differing buffer volumes in the first part of the protocol are just to achieve a standard 50ul starting point. Therefore if your sample volumes are 20ul, 30ul and 40ul, you will need to add 30ul, 20ul and 10ul respectively to bring total volumes to 50ul before proceeding with the kit reaction.



Although theoretically you can cut down all the volumes to half and still run the assay, we would not recommend doing it as it might end up compromising the efficiency of detection depending on the sample quality.


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

Read More

Answer

Thank you for your enquiry. Here are the details you requested. 1) Malate Assay Kit (ab83391): detects product formed (expected delivery date9th Jan) Step 1: Malate in sample + oxidation = product formed Step 2: Product formed + probe = color generation 2) Maltose Assay Kit (ab83388):detects product formed (no stock available, order lead time is 1-2 weeks) Step 1: Maltose in sample + alpha D glucosidase =two glucose units Step 2: Two glucose units + oxidation = product formed Step 3: Product formed + probe = color generation 3) Amylase Assay Kit (ab102523): direct detection/amount in sample(expected delivery date9th Jan) Step 1: Amylase in sample + substrate=fluorescence Note: Can be used in your research to measure enzyme activity over time! 4) Pyruvate Assay Kit (ab65342): detects product formed (expected delivery date9th Jan) Step 1: Pyruvatein sample + oxidation = product formed Step 2: Product formed + probe = color generation If there is anything else that I can help you with please let me know. Have a nice day!

Read More

Question

Happy New Year 2012 to you all .. I had a discussion with my boss today regarding the kits that you have offered. I would like to know following things.. · I understand that the kit is not able to directly measure the enzyme activity. However, we can correlate it back to enzyme activity by measuring color change, which is acceptable.. I would like to know what is this color change resulting from?? (i.e.) I would like to know what is the reaction that is happening for this color change formation… I would be interested to measure this color change, only if it is happening due to product formation….(i.e.) I won’t like to measure, if this color change is due to substrate consumption… The only reason behind this as follows: o For example, if the reaction is: known substrate+co-factor, buffers etcsample (containing enzyme to be tested)product § Now at time T0, if the substrate concentration is 100uM (for example), then the product concentration is Zero. § At time T1, if substrate concentration drops 95 uM, the product formed will be 5 uM ideally.. and so on for T2, T3… However, if we are evaluating for 5% change only, then generally if the kit is working based on substrate consumption, it is difficult to measure this difference significantly. Whereas, if the kit is working based on color change due to product formation, then that zero to 5 uM amount is generally found significant…. Hence, I would like to know on what method your kits are working… Please let me know if I was clear in explanation.. .I can arrange for the skype meet if there are any queries.. May I know how long does it take for the delivery to Singapore for the kits…? Thanks and regards

Read More
Answer

Thanks for contacting me. I have looked at these kits that we discussed and here are my thoughts: Malate Assay Kit (ab83391): This kit will only measure malate present in the sample. The enzyme used to produce the oxidized form of malate, the form that is detected by the probe in the kit, is considered proprietary by our supplying lab so unfortunately I can't provide any information regarding the oxidized form of malate. Maltose Assay Kit (ab83388): This kit could be used to measure alpha-D-glucosidase activity by measuring the conversion of maltose to 2 glucose molecules. Amylase Assay Kit (ab102523): This kit could be used to measure alpha-amylase activity by measuring the specific cleavage of ethylidene-pNP-G7 substrate. Pyruvate Assay Kit (ab65342): This kit could be used to measure pyruvate oxidase activity by measuring the oxidation of pyruvate present in the reaction. PicoProbe™ Acetyl CoA Assay Kit (ab87546): This kit can only be used to measure free acetyl CoA present in biological samples. I hope this is helpful. Please let me know if you need any further information.

Read More

Answer

Thank you for contacting us the other day. I have looked at the list of kits (attached file, summary that my colleague sent previously) that you are looking for and unfortunately, as my colleague has informed you, we only have 3 kits that may or may not be suitable for your research. 1) Malate dehydrogenase - ab83391 Malate Assay Kit In the assay, malate is specifically oxidized by using an enzyme mix to generate a product which reacts with WST substrate probe to generate color for detection. 2) Maltose hydrolase - ab83388 Maltose Assay Kit, ab65335 Maltose and Glucose Assay Kit The same theory applies here, where the glucose found in maltose, is used to produce a product that interacts with the glucose probe to generate color and fluorescence. 3) CoA synthetase - ab8756 PicoProbe™ Acetyl CoA Assay Kit Acetyl CoA is converted to CoA, then reacted to form NADH which interacts with PicoProbe to generate fluorescence. These kits are meant for detection of total amount of sample, NOT enzyme activity. However, the kits might be able to be used in reverse. Meaning that the enzyme in your soil sample will be used to convert a known amout of sample (standard) and later be quantified. But please bear in mind that this still gives you a reading in amount of sample detected but is correlated to the amount of enzyme in your soil sample. Other than the above suggestion, I am afraid that we do not have any kits that will suit your experiment. I have also taken the liberty of searching other possible kits available in the market using a Japanese website called Biohyakka but there are no specific kits found anywhere. (http://www.biohyakka.com/SearchResult.jsp) I hope the above information will be useful in some way. Please check with your supervisor if there are any other methods of enzyme detection normally used in your field of research. Have a nice day!

Read More

Answer

Thank you for your enquiry. Yes, you can use the cultured cells for ab83391. Here is the recommended sample preparation method for cultured cells; Cells (1-5X10e6) should be rapidly homogenized with two volumes of ice cold PBS or other buffer (pH 6.5-8). Enzymes in samples may interfere with the assay. We suggest deproteinizing your sample using a perchloric acid/KOH protocol or 10 kDa molecular weight cut off spin columns. Add 1-50 µl samples into duplicate wells of a 96-well plate and bring volume to 50 µl with Assay Buffer. We suggest testing several doses of your samples to ensure readings are within the standard curve range. I hope this information helps but please do not hesitate to contact me for additional information or assistance.

Read More

Answer

Thank you for your enquiry. I would be pleased to help answer your questions. 1. I can confirm you should be able to use these kits with blood samples. Generally serum or plasma that is obtained from blood samples is used for such purposes. However if you wish to use whole blood you will need to lyse the red blood cells in in each kits assay buffer and some optimization will be required by the customer in this case. 2. Overall you can use the following general procedure to prepare the sample. Samples can be deproteinized if indicated in respective data sheets either by using the PCA method or by using 10 kDa spin cut-off filters. For cell samples: i. Start with ~2X106 cells, suspend the cell pellet 500 μl (or ~4 volumes) of the assay buffer on ice. ii. Homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed, by viewing the cells under the microscope. iii. Spin down the sample and collect the supernatant. iv. Load the supernatant unto a 10 kda spin column for deproteinzation (please use this step only if indicated in the protocol provided with the online data sheet- this step is NOT required for enzyme assays). v. Use the eluate for the subsequent assays. Appropriate dilutions of the sample must be tested in order ensure the readings will fall within the linear range of the standard curve. 3. I am sorry to conrirm we do not sell the cut of columns or deprotinization kits. I would like to recommend checking the Biocompare website which has an excellent search facility that includes many suppliers. The links are: www.biocompare.com http://www.biocompare.com/ProductCategories/2045/Antibodies.html I hope this will be helpful to you. Should you have any further questions, please do not hesitate to contact us.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

登録