Human/Mouse/Rat/Pig ATPase, Na+/K+ Transporting Alpha 1 Polypeptide (ATP1a1) (Sandwich ELISA) ELISA Kit

Mfr.No.

SEG369Mi

Sensitivity

0.062ng/mL

Detection Range

0.156-10ng/mL

Reactivity

Human, Mouse, Rat, Pig

Assay Duration

Assay Type

Sandwich

Description

This assay has high sensitivity for detection of ATPase, Na+/K+ Transporting Alpha 1 Polypeptide (ATP1a1).
100% cross-reactivity of ATPase, Na+/K+ Transporting Alpha 1 Polypeptide (ATP1a1) was observed among Human, Mouse, Rat, Porcine.

Alternative Names

Sodium pump subunit alpha-1; Na(+)/K(+) ATPase alpha-1 subunit; Sodium/potassium-transporting ATPase subunit alpha-1

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 ATPase, Na+/K+ Transporting Alpha 1 Polypeptide (ATP1a1) and the recovery rates were calculated by comparing the measured value to the expected amount of ATPase, Na+/K+ Transporting Alpha 1 Polypeptide (ATP1a1) in samples.

Matrix	Recovery range (%)	Average(%)
serum(n=5)	80-90	87
EDTA plasma(n=5)	85-93	88
heparin plasma(n=5)	85-97	88

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of ATPase, Na+/K+ Transporting Alpha 1 Polypeptide (ATP1a1) 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)	82-96%	88-101%	89-103%	79-103%
EDTA plasma(n=5)	88-96%	86-94%	89-103%	80-98%
heparin plasma(n=5)	95-105%	87-95%	78-92%	82-97%

Features

Detects protein (regular version)

Species

Human///Mouse///Rat///Pig

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 ATPase, Na+/K+ Transporting Alpha 1 Polypeptide (ATP1a1) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level ATPase, Na+/K+ Transporting Alpha 1 Polypeptide (ATP1a1) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: C<10%
Inter-Assay: C<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 ATPase, Na+/K+ Transporting Alpha 1 Polypeptide (ATP1a1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to ATPase, Na+/K+ Transporting Alpha 1 Polypeptide (ATP1a1). 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 ATPase, Na+/K+ Transporting Alpha 1 Polypeptide (ATP1a1), 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 ATPase, Na+/K+ Transporting Alpha 1 Polypeptide (ATP1a1) 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	476
UniProt ID	P05023

Applications	ELISA
Research Area	Signal transduction

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

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ELISA Kit for ATPase, Na+/K+ Transporting Alpha 1 Polypeptide (ATP1a1) (Human/Mouse/Rat/Pig)

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