Human anti-HSPA5 (anti-Heat shock protein family A member 5) ELISA Kit

Mfr.No.

EH4919

Sensitivity

0.234ng/ml

Detection Range

0.391-25ng/ml

Reactivity

Human

Assay Duration

3 hours

Assay Type

Indirect

Description

This kit is based on indirect ELISA detection method and takes 3h assay time. The microplate provided in this kit has been precoated with antigen. HRP-antibody is used as detection antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add HRP-detection antibody, then it binds with HSPA5-Ab bound to precoated antibody. Wash unbound components and 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. Calculate the concentration of HSPA5-Ab in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.

Alternative Names

HSPA5 antibody ELISA Kit, Heat shock protein family A member 5 antibody ELISA Kit, Endoplasmic reticulum chaperone BiP antibody ELISA Kit

Recovery

Add a certain amount of HSPA5-Ab into the sample. Calculate the recovery by comparing the measured value with the expected amount of HSPA5-Ab in the sample.

Sample Type	Recovery Range(%)	Average(%)
serum(n=10)	91-102	99
EDTA plasma(n=10)	90-105	97
Heparin plasma(n=10)	85-102	92

Linearity

Dilute the sample with a certain amount of HSPA5-Ab at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type	1:2	1:4	1:8
serum(n=10)	86-103%	85-105%	94-101%
EDTA plasma(n=10)	92-100%	83-101%	83-96%
Heparin plasma(n=10)	82-96%	85-98%	87-100%

Specificity

Specifically binds with HSPA5-Ab , 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.(ng/ml)	OD-1	OD-2	Average
0	0.088	0.09	0.089
0.391	0.197	0.203	0.2
0.781	0.295	0.303	0.299
1.562	0.468	0.482	0.475
3.125	0.953	0.981	0.967
6.25	1.312	1.35	1.331
12.5	1.762	1.814	1.788
25	2.15	2.212	2.181

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 (ng/ml)	0.77	2.94	11.81	0.75	3.17	12.58
Standard deviation	0.03	0.17	0.58	0.04	0.15	0.4
CV(%)	3.25	5.86	4.95	5.43	4.68	3.19

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	P11021

Applications	ELISA

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

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SKU	Supplier No.	Size	Your price	Quantity
BHE10807258	EH4919	48T	$280		Add to Cart
BHE10807258	EH4919	96T	$400		Add to Cart

Human anti-HSPA5 (anti-Heat shock protein family A member 5) ELISA Kit

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