Rat Creatine Kinase, Muscle (CKM) (Sandwich ELISA) ELISA Kit

Mfr.No.

SEA109Ra

Sensitivity

0.61ng/mL

Detection Range

1.56-100ng/mL

Reactivity

Rat

Sample Type

Cell lysates, Cell culture supernates, Serum, Tissue homogenates, Plasma, Other biological fluids

Assay Duration

Assay Type

Sandwich

Description

This assay has high sensitivity and excellent specificity for detection of Creatine Kinase, Muscle (CKM).
No significant cross-reactivity or interference between Creatine Kinase, Muscle (CKM) and analogues was observed.

Alternative Names

CK-MM; CKMM; CK3; CK-M; MM-CK; M-CK; Creatine kinase M-type, N-terminally processed

Detection Method

Colormetric

Kit Components

Reagents	Quantity	Reagents	Quantity
Pre-coated, ready to use 96-well strip plate	1	Plate sealer for 96 wells	4
Standard	2	Standard Diluent	1×20mL
Detection Reagent A	1×120µL	Assay Diluent A	1×12mL
Detection Reagent B	1×120µL	Assay Diluent B	1×12mL
TMB Substrate	1×9mL	Stop Solution	1×6mL
Wash Buffer (30 × concentrate)	1×20mL	Instruction manual	1

Recovery

Matrices listed below were spiked with certain level of recombinant Creatine Kinase, Muscle (CKM) and the recovery rates were calculated by comparing the measured value to the expected amount of Creatine Kinase, Muscle (CKM) in samples.

Matrix	Recovery range (%)	Average(%)
serum(n=5)	81-104	89
EDTA plasma(n=5)	81-101	93
heparin plasma(n=5)	87-103	94

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Creatine Kinase, Muscle (CKM) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample	1:2	1:4	1:8	1:16
serum(n=5)	89-105%	78-93%	80-103%	79-93%
EDTA plasma(n=5)	85-98%	89-103%	99-105%	99-105%
heparin plasma(n=5)	87-103%	96-104%	79-96%	81-90%

Features

Detects protein (regular version)

Species

Rat

Precautions

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Creatine Kinase, Muscle (CKM) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Creatine Kinase, Muscle (CKM) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Notes

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Creatine Kinase, Muscle (CKM). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Creatine Kinase, Muscle (CKM). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Creatine Kinase, Muscle (CKM), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Creatine Kinase, Muscle (CKM) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Usage Notes

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

Protocol

Datasheet

Gene ID	24265
UniProt ID	P00564
Antigen	CKM

Applications	ELISA
Research Area	Enzyme///Kinase

Storage	4°C for 1 month/ -20°C for long-term(within shelf life)
Shelf Life	one year

Reference	Extraocular Muscle Characteristics Related to Myasthenia Gravis Susceptibility

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ELISA Kit for Creatine Kinase, Muscle (CKM) (Rat)

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