Human PRKAA1 (5 -AMP-activated protein kinase catalytic subunit alpha-1) ELISA Kit

Mfr.No.

EH2503

Sensitivity

46.875pg/ml

Detection Range

78.125-5000pg/ml

Reactivity

Human

Assay Duration

2 hours

Assay Type

Competitive

Description

This kit is based on Competitive-ELISA detection method and takes 2h assay time. The microplate provided in this kit has been precoated with PRKAA1. PRKAA1 in the sample or standard competes with a fixed amount of biotin-labeled PRKAA1 for sites on a pre-coated antibody specific to PRKAA1. Free components are washed off. After adding HRP-Streptavidin Conjugate (SABC), biotin specifically binds with streptavidin to form immune complexes. Wash off unbound conjugate. Add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Compare OD450 value of sample with the standard curve through curve fitting software to determine the concentration of PRKAA1 in the sample. The concentration of the target substance is inversely proportional to the OD450 value.

Alternative Names

5'-AMP-activated protein kinase catalytic subunit alpha-1 ELISA Kit, AMPK subunit alpha-1 ELISA Kit, Acetyl-CoA carboxylase kinase ELISA Kit, ACACA kinase ELISA Kit, Hydroxymethylglutaryl-CoA reductase kinase ELISA Kit, HMGCR kinase ELISA Kit, Tau-protein kinase PRKAA1 ELISA Kit, PRKAA1 ELISA Kit, AMPK1 ELISA Kit

Detection Method

Coated with Antigen

Kit Components

Kit Components	Item	Size(48T)	Size(96T)	Storage Condition for Opened Kit
E001	ELISA Microplate(Dismountable)	8x6	8x12	Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E002	Lyophilized Standard	1vial	2vial	Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E003	Biotin-labeled Antibody(Concentrated, 100X)	30ul	60ul	2-8°C (Avoid Direct Light)
E034	HRP-Streptavidin Conjugate(SABC, 100X)	60ul	120ul
E024	TMB Substrate	5ml	10ml
E039	Sample Dilution Buffer	10ml	20ml	2-8°C
E040	Antibody Dilution Buffer	5ml	10ml
E049	SABC Dilution Buffer	5ml	10ml
E026	Stop Solution	5ml	10ml
E038	Wash Buffer(Concentrated, 25X)	15ml	30ml
E006	Plate Sealer	3 pieces	5 pieces
E007	Product Description	1 copy	1 copy

Recovery

Add a certain amount of PRKAA1 into the sample. Calculate the recovery by comparing the measured value with the expected amount of PRKAA1 in the sample.

Sample Type	Recovery Range(%)	Average(%)
serum(n=10)	93-104	99
EDTA plasma(n=10)	86-105	96
Heparin plasma(n=10)	87-104	93

Linearity

Dilute the sample with a certain amount of PRKAA1 at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type	1:2	1:4	1:8
serum(n=10)	86-102%	85-102%	88-101%
EDTA plasma(n=10)	85-101%	93-105%	82-97%
Heparin plasma(n=10)	88-97%	82-99%	83-99%

Specificity

Specifically binds with PRKAA1 , no obvious cross reaction with other analogues.

Standard Curve

This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.) Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.

STD.(pg/ml)	OD-1	OD-2	Average
0	2.396	2.519	2.445
78.125	1.901	1.998	1.94
156.25	1.672	1.758	1.707
312.5	1.596	1.678	1.629
625	1.229	1.292	1.254
1250	0.878	0.923	0.896
2500	0.659	0.692	0.672
5000	0.37	0.389	0.377

Sample Type

Tissue lysate, Cell lysates, Cell culture supernatants, Plasma, Other biological fluids

Components

Microplate,Lyophilized Standard,buffer,SABC,Label antibody,SABC,TMB,stop solution

Precision

Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.
Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.

Item	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/ml)	153.9	594.8	2456	157.9	616.8	2453
Standard deviation	8.14	30.75	125.5	8.02	30.96	130.01
CV(%)	5.29	5.17	5.11	5.08	5.02	5.3

Background

PRKAA1/2.7.11.26/Tau-protein kinase PRKAA1)/2.7.11.27/Tau-protein kinase PRKAA1(ACACA kinase)/2.7.11.31/Hydroxymethylglutaryl-CoA reductase kinase(HMGCR kinase)/AMPK1/AMPK subunit alpha-1

Stability

Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.

ELISA kit(n=5)	37°C for 1 month	2-8°C for 6 months
Average(%)	80	95-100

Notes

For Research Use Only

Usage Notes

Step 1:
Take out the required plate wells, add 50ul Cap/Det Ab into each well, then add 50ul Standard or Sample into individual well. (When adding standard or sample, the disposable tip lightly touches the liquid level. Change the disposable tips for different samples and standards.) Gently tap the plate for 10s to ensure thorough mixing then static incubate for 60 minutes at 37°C.

Washing:
Wash the plate twice without immersion.

Step 2:
Add 100ul HRP-Streptavidin (orange) into each well, seal the plate and static incubate for 30 minutes at 37°C.

Washing:
Wash the plate five times without immersion.

Step 3:
Add 90ul TMB substrate solution, seal the plate and static incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)

Step 4:
Add 50ul stop solution. Read at 450nm immediately and calculate.

UniProt ID	Q13131

Applications	ELISA
Applications Notes	PRKAA1 ELISA Kit allows for the in vitro quantitative determination of PRKAA1 concentrations in serum, plasma, tissue homogenates and other biological fluids.

Storage	2-8°C(Sealed), Don't cryopreserve.

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SKU	Supplier No.	Size	Your price	Quantity
BHE10805469	EH2503	48T	$280		Add to Cart
BHE10805469	EH2503	96T	$400		Add to Cart

Human PRKAA1 (5 -AMP-activated protein kinase catalytic subunit alpha-1) ELISA Kit

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