Multiplex Assay Kit for Heat Shock 70kDa Protein 5 (HSPA5) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Heat Shock 70kDa Protein 5 (HSPA5) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Heat Shock 70kDa Protein 5 (HSPA5) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-101 88
EDTA plasma(n=5) 82-99 85
heparin plasma(n=5) 99-105 102
sodium citrate plasma(n=5) 80-101 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Heat Shock 70kDa Protein 5 (HSPA5) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 95-104% 97-104% 89-101% 78-101%
EDTA plasma(n=5) 98-105% 84-96% 88-102% 85-99%
heparin plasma(n=5) 89-97% 81-105% 94-104% 94-102%
sodium citrate plasma(n=5) 94-105% 88-102% 93-105% 84-104%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:HSPA5) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

GIVEAWAYS

INCREMENT SERVICES

Magazine Citations
41 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
pharmacological research Neuroprotection in non-transgenic and transgenic mouse models of Alzheimer's disease by positive modulation of σ1 receptors Pubmed: 31048034
EPILEPSY & BEHAVIOR Fenfluramine acts as a positive modulator of sigma-1 receptors Pubmed: 32169824
Journal of Oncology Using Serological Proteome Analysis to Identify and Evaluate Anti-GRP78 Autoantibody as Biomarker in the Detection of Gastric Cancer 33381181
Heliyon Glucose-regulated protein 78 (GRP78) in renal cell carcinoma: A novel biomarker for predicting tumor behavior
Medicine (Baltimore) Levels of circulating GRP78 and CHOP in endoplasmic reticulum stress pathways in Chinese type 2 diabetic kidney disease patients 34414940
Catalog No. Related products for research use of Rattus norvegicus (Rat) Organism species Applications (RESEARCH USE ONLY!)
RPC343Ra01 Recombinant Heat Shock 70kDa Protein 5 (HSPA5) Positive Control; Immunogen; SDS-PAGE; WB.
PAC343Ra01 Polyclonal Antibody to Heat Shock 70kDa Protein 5 (HSPA5) WB; IHC; ICC; IP.
MAC343Ra21 Monoclonal Antibody to Heat Shock 70kDa Protein 5 (HSPA5) WB; IHC; ICC; IP.
SEC343Ra ELISA Kit for Heat Shock 70kDa Protein 5 (HSPA5) Enzyme-linked immunosorbent assay for Antigen Detection.
LMC343Ra Multiplex Assay Kit for Heat Shock 70kDa Protein 5 (HSPA5) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.
KSC343Ra01 ELISA Kit DIY Materials for Heat Shock 70kDa Protein 5 (HSPA5) Main materials for "Do It (ELISA Kit) Yourself".