Rat HSP-70/HSPA9(Heat Shock Protein 70) ELISA Kit

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Product No

FY-ER5128

Sample Type

Serum, Plasma, Tissue homogenate and Other biological samples

Assay Time

3.5H

Price($)

280 400

Size

48T 96T

Purchase details

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DataSheet

说明书 MSDS

MSDS

Product Details

Product Name in English:

Rat HSP-70/HSPA9(Heat Shock Protein 70) ELISA Kit

Species:

Rat

Alternative Name:

HSPA9, CSA, GRP-75, GRP75, HSPA9B, MOT, MOT2, MTHSP75, PBP74

Uniport ID:

P48721

Assay Type:

Sandwich

Standard:

20ng/mL

Sensitivity:

0.19 ng/mL

Detection Range:

0.32-20ng/mL

Assay Time:

3.5H

Sample Type:

Serum, Plasma, Tissue homogenate and Other biological samples

Research:

Developmental science;

Test Principle:

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 HSP-70/HSPA9. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to HSP-70/HSPA9. 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 HSP-70/HSPA9, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of HSP-70/HSPA9 in the samples is then determined by comparing the OD of the samples to the standard curve.

Technical Data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration	OD1	OD2	Average OD	Corrected
20	2.334	2.341	2.338	2.284
10	1.54	1.555	1.548	1.49
5	0.909	0.916	0.913	0.859
2.5	0.426	0.426	0.426	0.376
1.25	0.247	0.249	0.248	0.197
0.63	0.147	0.147	0.147	0.097
0.32	0.1	0.101	0.101	0.05
0	0.05	0.05	0.05	0

Precision

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

Item	Intra-assay Precision	Intra-assay Precision
Sample	3	3
Replicate	9	18
CV(%)	5	8

Rate of recovery

The recovery of 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)	82-95	88
EDTA plasma (n=8)	88-102	95
Cell culture media (n=8)	80-95	87

Linear

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

		Serum (n=5)	EDTA plasma (n=5)	Cell culture media (n=5)
1:2	Range (%)	93-106	82-96	93-105
1:2	Average (%)	101	84	103
1:4	Range (%)	83-95	86-98	87-103
1:4	Average (%)	93	96	87
1:8	Range (%)	88-103	93-101	85-103
1:8	Average (%)	94	96	95
1:16	Range (%)	83-97	82-101	82-96
1:16	Average (%)	87	98	88

Assay Procedures

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