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.
Fortfahren mit
MAK116
Kreatinkinase-Aktivitäts-Assay-Kit
sufficient for 100 tests (Ultraviolet)
Synonym(e):
CK-Aktivitäts-Assay-Kit, Kreatin-Phosphokinase-Aktivitäts-Assay-Kit
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| Größe/SKU | Verfügbarkeit | Preis |
|---|---|---|
1 kit | Warenkorb auf Verfügbarkeit prüfen | € 587,00 |
Über diesen Artikel
NACRES:
NA.84
UNSPSC Code:
12161503
Technischer Dienst
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Unterstützung erhaltenusage
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
Die Kreatinkinase (CK), auch Phosphokreatinkinase genannt, ist ein Enzym, das die Übertragung einer Phosphatgruppe von ATP auf Kreatin katalysiert und dabei Phosphokreatin, ein wichtiges Energiereservoir im Muskel- und Gehirngewebe, erzeugt. CK ist ein dimeres Protein, das aus den Untereinheiten B („brain“/Gehirn) und M (Muskel) besteht. Es wurden drei Isoenzyme beobachtet (CK-MM, CK-MB und CK-BB). Die CK-Werte sind bei verschiedenen pathologischen Zuständen wie Herzinfarkt, Rhabdomyolyse, Muskeldystrophie und Nierenversagen erhöht.
Application
Geeignet zum Nachweis der Kreatinkinase-Aktivität in einer Vielzahl von Proben wie Serum und Plasma. Mit diesem Kit können die Auswirkungen verschiedener Verbindungen auf die Kreatinkinase-Aktivität untersucht werden.
Biochem/physiol Actions
Die Kreatinkinase-Aktivität wird durch eine gekoppelte Enzymreaktion bestimmt, die zur Produktion von NADPH führt, das bei 340 nm gemessen wird und proportional zur CK-Aktivität in der Probe ist. Bei dieser Reaktion werden Phosphokreatin und ADP in Kreatin und ATP umgewandelt. Das erzeugte ATP wird von der Hexokinase zur Phosphorylierung von Glucose verwendet, wodurch Glucose-6-Phosphat entsteht, das von NADP in Gegenwart von Glucose-6-Phosphat-Dehydrogenase zu NADPH und 6-Phospho-D-Gluconat oxidiert wird. Eine CK-Einheit ist als die Enzymmenge definiert, die bei einem pH-Wert von 6,0 pro Minute 1,0 μMol Phosphat von Phosphokreatin auf ADP überträgt. Dieses Kit hat einen linearen Bereich von 30–1.800 Einheiten/L CK-Aktivität.
Features and Benefits
Kompatibel mit Handhabungssystemen mit hohem Durchsatz.
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Dieser Artikel | ||
|---|---|---|
| description sufficient for 100 tests (Ultraviolet) | description sufficient for 100 colorimetric or fluorometric tests | description for serum, plasma, cell culture supernatants and urine |
| Gene Information human ... CKB(1152), CKM(1158) | Gene Information - | Gene Information human ... CKB(1152), CKM(1158) |
| usage sufficient for 100 tests (Ultraviolet) | usage sufficient for 100 colorimetric or fluorometric tests | usage - |
| detection method colorimetric | detection method colorimetric, fluorometric | detection method colorimetric |
| shipped in dry ice | shipped in dry ice | shipped in wet ice |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
| relevant disease(s) cardiovascular diseases; genitourinary disorders; aging/geriatric diseases; orthopedic diseases | relevant disease(s) - | relevant disease(s) - |
signalword
Danger
hcodes
pcodes
Hazard Classifications
Resp. Sens. 1
Lagerklasse
12 - Non Combustible Liquids
wgk
WGK 3
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How much mouse blood/plasma would you recommend using for successful measurement?
1 answer-
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How can I determine the shelf life / expiration / retest date of this product?
1 answer-
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?
1 answer-
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?
1 answer-
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?
1 answer-
Yes, this kit is suitable for determining CK levels in mammalian samples.
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