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Human PKM2(Pyruvate Kinase, Muscle) ELISA Kit

Citations(5)Uniprot : P14618

Cat.No.:E-EL-H6063
Reactivity: Human

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Product Details

Properties

Assay type	Sandwich-ELISA
Format	96T/48T/24
Assay time	3.5h
Detection range	15.63-1000 IU/mL
Sensitivity	9.38 IU/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 100μL
Specificity	This kit recognizes Human PKM2 in samples. No significant cross-reactivity or interference between Human PKM2 and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of Human PKM2 concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human PKM2. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human PKM2 and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human PKM2, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human PKM2. You can calculate the concentration of Human PKM2 in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 12 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 12 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°C, 12 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25×)	1 vial, 30 mL
Substrate Reagent	1 vial, 10 mL	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8°C
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PKM2 were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PKM2 were tested on 3 different plates, 20 replicates in each plate.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (IU/mL)	47.47	116.17	471.44	47.83	121.50	409.39
Standard deviation	2.69	5.66	22.06	2.50	5.67	17.19
CV (%)	5.67	4.87	4.68	5.23	4.67	4.20

Recovery

The recovery of Human PKM2 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type	Range (%)	Average Recovery (%)
Serum(n=8)	88-100	95
EDTA plasma (n=8)	98-110	105
Cell culture media (n=8)	90-99	94

Linearity

Samples were spiked with high concentrations of Human PKM2 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database Links	SwissProt: P14618
Research Area	Cancer, Epigenetics and Nuclear Signaling, Metabolism, Signal transduction

Assay Procedures

	1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
	2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
	3. Aspirate and wash the plate for 3 times
	4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	6. Add 50μL Stop Solution
	7. Read the plate at 450nm immediately. Calculation of the results

Citations

CLINICAL NEUROLOGY AND NEUROSURGERY (2023) IF: 1.885

Negative correlation between serum pyruvate kinase M2 and cognitive function in patients with cerebral small vessel disease

DOI: 10.1016/j.clineuro.2023.107586
	Sample: Serum

CYTOTECHNOLOGY (2021) IF: 2.058

Investigation of glucose catabolism in hypoxic Mcf 7 breast cancer culture

DOI: 10.1007/s10616-021-00459-2
PMID: 33927477	Sample: Cell（Mcf-7 Cancer Cells）

CLINICAL BIOCHEMISTRY (2017) IF: 2.434

Elevation of serum pyruvate kinase M2 (PKM2) in IBD and its relationship to IBD indices

DOI: 10.1016/j.clinbiochem.2017.12.007
	Sample: Serum,Cell Lysate

JOURNAL OF INTERFERON AND CYTOKINE RESEARCH (2022) IF: 3.657

Can the Prognosis of COVID-19 Disease Be Determined by Fecal Markers and Cytokines?

DOI: 10.1089/jir.2022.0098
PMID: 36179036	Sample: Fecal

Toxics (2023) IF: 4.472

A Proposed Association between Improving Energy Metabolism of HepG2 Cells by Plant Extracts and Increasing Their Sensitivity to Doxorubicin

DOI: 10.3390/toxics11020182
	Sample: Hepg2 Cell

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Reviews/Q&A

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Reviews
Q&A

c******************mSubmitted [ Jun 23 2022 ]

Asked: How does the IU/mL measurement translate to actual protein concentration in pg/mL or ng/mL?

adminSubmitted [ Jun 23 2022 ]

Answered: Sorry, there is no international conversion factor for these two units, the units couldn't be converted from IU/mL to pg/mL or ng/mL.

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