Bovine/Goat/Sheep Heat Shock Protein 70 (HSP70) (Sandwich ELISA) ELISA Kit

Mfr.No.

SEA873Mi

Description

This assay has high sensitivity for detection of Heat Shock Protein 70 (HSP70).
100% cross-reactivity of Heat Shock Protein 70 (HSP70) was observed among Bovine, Caprine, Ovine.

Alternative Names

DnaK; 70 Kilodalton Heat Shock Proteins

Reactivity

Bovine, Goat, Sheep

Detection Method

Colormetric

Detection Range

3.12-200ng/mL

Sample Type

Other biological fluids, Serum, Plasma

Assay Type

Sandwich

Assay Duration

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 Heat Shock Protein 70 (HSP70) and the recovery rates were calculated by comparing the measured value to the expected amount of Heat Shock Protein 70 (HSP70) in samples.

Matrix	Recovery range (%)	Average(%)
serum(n=5)	82-103	91
EDTA plasma(n=5)	80-101	85
heparin plasma(n=5)	91-105	98

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Heat Shock Protein 70 (HSP70) 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)	96-105%	78-95%	89-96%	79-101%
EDTA plasma(n=5)	90-104%	95-102%	94-103%	78-103%
heparin plasma(n=5)	86-94%	78-98%	89-103%	82-99%

Features

Detects protein (regular version)

Species

Bovine///Goat///Sheep

Sensitivity

1.31ng/mL

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.

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 Heat Shock Protein 70 (HSP70). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Heat Shock Protein 70 (HSP70). 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 Heat Shock Protein 70 (HSP70), 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 Heat Shock Protein 70 (HSP70) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Antigen	HSP70
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.

Applications	ELISA
Research Area	Tumor immunity
Protocol	Datasheet
Precision	Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Heat Shock Protein 70 (HSP70) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Heat Shock Protein 70 (HSP70) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 Intra-Assay: C<10% Inter-Assay: C<12%

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

Reference	The effect of heat stress on gene expression and synthesis of heat‐shock and milk proteins in bovine mammary epithelial cells

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SKU	Supplier No.	Size	Your price
BHE10103021	SEA873Mi	96T*1	$648	Add to Cart
BHE10103021	SEA873Mi	96T*5	$2916	Add to Cart
BHE10103021	SEA873Mi	96T*10	$5508	Add to Cart

ELISA Kit for Heat Shock Protein 70 (HSP70) (Bovine/Goat/Sheep)

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