ELISA Kit for Heat Shock 70kDa Protein 5 (HSPA5) Homo sapiens (Human) Sandwich ELISA

MIF2; BIP; GRP78; Immunoglobulin heavy chain-binding protein; Glucose Regulated Protein 78; Binding Immunoglobulin Protein; Endoplasmic reticulum lumenal Ca binding grp78

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  • ELISA Kit for Heat Shock 70kDa Protein 5 (HSPA5) Packages (Simulation)
  • ELISA Kit for Heat Shock 70kDa Protein 5 (HSPA5) Packages (Simulation)
  • ELISA Kit for Heat Shock 70kDa Protein 5 (HSPA5) Results demonstration
  • SEC343Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Heat Shock 70kDa Protein 5 (HSPA5) and the recovery rates were calculated by comparing the measured value to the expected amount of Heat Shock 70kDa Protein 5 (HSPA5) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 83-95 91
EDTA plasma(n=5) 95-103 98
heparin plasma(n=5) 90-99 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Heat Shock 70kDa Protein 5 (HSPA5) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Heat Shock 70kDa Protein 5 (HSPA5) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Heat Shock 70kDa Protein 5 (HSPA5) 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) 82-99% 96-104% 78-101% 97-104%
EDTA plasma(n=5) 78-92% 93-101% 89-103% 86-101%
heparin plasma(n=5) 85-96% 80-92% 93-102% 84-101%

Stability

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.

Reagents and materials provided

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

Assay procedure summary

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.

ELISA Kit for Heat Shock 70kDa Protein 5 (HSPA5)

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 Heat Shock 70kDa Protein 5 (HSPA5). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Heat Shock 70kDa Protein 5 (HSPA5). 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 70kDa Protein 5 (HSPA5), 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 70kDa Protein 5 (HSPA5) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Deficiency of Rac1 Blocks NADPH Oxidase Activation, Inhibits Endoplasmic Reticulum Stress, and Reduces Myocardial Remodeling in a Mouse Model of Type 1 Diabetes.PubMed: 20522592
  • Neuroprotection in non-transgenic and transgenic mouse models of Alzheimer's disease by positive modulation of σ1 receptorsPubmed: 31048034
  • Fenfluramine acts as a positive modulator of sigma-1 receptorsPubmed: 32169824
  • Using Serological Proteome Analysis to Identify and Evaluate Anti-GRP78 Autoantibody as Biomarker in the Detection of Gastric Cancer33381181
  • Glucose-regulated protein 78 (GRP78) in renal cell carcinoma: A novel biomarker for predicting tumor behavior
  • Levels of circulating GRP78 and CHOP in endoplasmic reticulum stress pathways in Chinese type 2 diabetic kidney disease patients34414940

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