It is recommended to refer to the protocol for appropriate usage and sample amounts. According to step 3 in the Assay reaction section, transferring 10 µL of samples into separate wells is advised.
MAK116
Creatine Kinase Activity Assay Kit
sufficient for 100 tests (Ultraviolet)
Creatine Kinase Activity Assay Kit
Synonym(s):
CK Activity Assay Kit, Creatine Phosphokinase Activity Assay Kit
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| Size/SKU | Availability | Price |
|---|---|---|
1 kit | Estimated to ship onAugust 12, 2026fromSAINT LOUIS | $670.00 |
About This Item
NACRES:
NA.84
UNSPSC Code:
12161503
usage
sufficient for 100 tests (Ultraviolet)
detection method
colorimetric
relevant disease(s)
cardiovascular diseases; genitourinary disorders; aging/geriatric diseases; orthopedic diseases
shipped in
dry ice
storage temp.
−20°C
Gene Information
human ... CKB(1152), CKM(1158)
mouse ... CKB(12709), CKM(12715)
rat ... CKB(24264), CKM(24265)
General description
Creatine Kinase (CK), also known as phosphocreatine kinase, is an enzyme that catalyzes the transfer of one phosphate group from ATP to creatine generating phosphocreatine, an important energy reservoir in muscle and brain tissue. CK is a dimeric protein made up of B (brain) and M (muscle) subunits. Three isoenzymes, CK-MM, CK-MB, and CK-BB, have been observed. CK levels are elevated in various pathological conditions including myocardial infarction, rhabdomyolysis, muscular dystrophy, and renal failure.
Application
Suitable for the detection of Creatine Kinase activity in a variety of samples such as serum and plasma. This kit can be used to study the effects of various compounds on Creatine Kinase activity.
Biochem/physiol Actions
Creatine Kinase activity is determined by a coupled enzyme reaction resulting in the production of NADPH, measured at 340 nm, proportionate to the CK activity present in the sample. In this reaction, phosphocreatine and ADP are converted to creatine and ATP. The generated ATP is used by hexokinase to phosphorylate glucose resulting in glucose-6-phosphate, which is oxidized by NADP in the presence of glucose-6-phosphate dehydrogenase to produce NADPH and 6-phospho-D-gluconate. One unit of CK is the amount of enzyme that will transfer 1.0 μmole of phosphate from phosphocreatine to ADP per minute at pH 6.0. This kit has a linear range of 30–1,800 units/L CK activity.
Features and Benefits
Compatible with high-throughput handling systems.
1 of 1
This Item | |||
|---|---|---|---|
| description sufficient for 100 tests (Ultraviolet) | description sufficient for 100 colorimetric or fluorometric tests | description for serum, plasma, cell culture supernatants and urine | description sufficient for 100 tests (UV) |
| Gene Information human ... CKB(1152), CKM(1158) | Gene Information - | Gene Information human ... CKB(1152), CKM(1158) | Gene Information human ... GLO1(2739) |
| usage sufficient for 100 tests (Ultraviolet) | usage sufficient for 100 colorimetric or fluorometric tests | usage - | usage sufficient for 100 tests (UV) |
| detection method colorimetric | detection method colorimetric, fluorometric | detection method colorimetric | detection method colorimetric |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
| shipped in dry ice | shipped in dry ice | shipped in wet ice | shipped in - |
| relevant disease(s) cardiovascular diseases; genitourinary disorders; aging/geriatric diseases; orthopedic diseases | relevant disease(s) - | relevant disease(s) - | relevant disease(s) cancer |
signalword
Danger
hcodes
pcodes
Hazard Classifications
Resp. Sens. 1
Storage Class
12 - Non Combustible Liquids
wgk
WGK 3
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How much mouse blood/plasma would you recommend using for successful measurement?
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How can I determine the shelf life / expiration / retest date of this product?
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If this product has an expiration or retest date, it will be shown on the Certificate of Analysis (COA, CofA). If there is no retest or expiration date listed on the product's COA, we do not have suitable stability data to determine a shelf life. For these products, the only date on the COA will be the release date; a retest, expiration, or use-by-date will not be displayed.
For all products, we recommend handling per defined conditions as printed in our product literature and website product descriptions. We recommend that products should be routinely inspected by customers to ensure they perform as expected.
For products without retest or expiration dates, our standard warranty of 1 year from the date of shipment is applicable.
For more information, please refer to the Product Dating Information document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/418/501/product-dating-information-06-25-mk.pdfHelpful?
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How is shipping temperature determined? And how is it related to the product storage temperature?
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Products may be shipped at a different temperature than the recommended long-term storage temperature. If the product quality is sensitive to short-term exposure to conditions other than the recommended long-term storage, it will be shipped on wet or dry-ice. If the product quality is NOT affected by short-term exposure to conditions other than the recommended long-term storage, it will be shipped at ambient temperature. As shipping routes are configured for minimum transit times, shipping at ambient temperature helps control shipping costs for our customers. For more information, please refer to the Storage and Transport Conditions document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/316/622/storage-transport-conditions-mk.pdf
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How long are the provided solutions stable for when stored at -20C? Does the assay lose efficacy through repeated freeze/thaw cycles of the assay solutions?
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The unopened kit is expiration dated on a batch-by-batch basis and the certificate of analysis for a specific lot would need to be reviewed for this information. Please review the "product documentation" section of the website to pull a sample CoA or to input the lot number for the received lot.
https://www.sigmaaldrich.com/product/sigma/MAK116#product-documentation
There is no data available on the amount of freeze/thaws this kit can withstand. It is expected that repeated freeze/thaws of enzyme and substrate solution could affect the activity. Therefore, it would be advised to aliquot multiple small volumes of the kit reagents for one-time use to avoid repeated freeze/thaw cycles.
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What is the required cell count for this procedure, and at what cell concentration is the supernatant obtained?
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For preparing cell samples for use with MAK116, the Creatine Kinase Activity Assay Kit, the vendor recommends using the cell lysate for the assay. The number of cells to use will depend on the CK activity of the cells, and the final CK activity of the sample used for the assay should be within 5 to 300 U/L creatine kinase (the linear detection range in a 96-well plate assay). Here's an example of a sample preparation protocol for adherent or suspension cells:
1. Harvest the required number of cells for each assay, with an initial recommendation of two million or 2 x 10^6 cells.
2. Wash the cells with cold PBS and resuspend them in 400 µL PBS on ice.
3. Lyse the cells by homogenization (10-20 passes in a Dounce homogenizer on ice) or by sonication (preferably performed in an ice-water bath). The degree of cell lysis can be checked under a microscope.
4. Centrifuge the homogenate at 14,000 g for 10 minutes and transfer the clear supernatant into a clean tube.
5. Conduct a pilot test of the sample at different dilutions. Choose a dilution with readings within the range of the calibrator for further assays. Most samples can be stored at -80°C if not assayed immediately.Helpful?
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Can this kit be used on an adherent cell line (rat species)?
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Creatine kinase is generally found in muscle tissue or in plasma. The MAK116 has been published in the scientific literature for samples such as rat serum, plasma and tissue. There are not any articles referring specifically to rat adherent cell lines. Further research and development for this kit would be necessary to validate for this application. The insert does provide instructions for working with adherent cell lysate.
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/355/268/mak116bul.pdfHelpful?
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Regarding the creatine kinase assay, will this detect CK activity from other mammalian species such as bovine?
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Yes, this kit is suitable for determining CK levels in mammalian samples.
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