Multiplex Assay Kit for Heat Shock Protein 27 (Hsp27) ,etc. by FLIA (Flow Luminescence Immunoassay) Bos taurus; Bovine (Cattle) Multiplex ELISA

HSPB1; HSP-B1; CMT2F; HSP28; Hsp25; Heat Shock 27kDa Protein 1; 28 kDa heat shock protein; Heat shock protein beta-1; Estrogen-regulated 24 kDa protein;

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

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  • Multiplex Assay Kit for Heat Shock Protein 27 (Hsp27) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Heat Shock Protein 27 (Hsp27) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Heat Shock Protein 27 (Hsp27) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA693Bo.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Heat Shock Protein 27 (Hsp27) and the recovery rates were calculated by comparing the measured value to the expected amount of Heat Shock Protein 27 (Hsp27) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 93-104 98
EDTA plasma(n=5) 92-99 96
heparin plasma(n=5) 93-101 97

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Heat Shock Protein 27 (Hsp27) 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 27 (Hsp27) 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 Protein 27 (Hsp27) 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) 79-91% 97-105% 81-99% 79-101%
EDTA plasma(n=5) 87-101% 92-99% 78-97% 80-93%
heparin plasma(n=5) 96-103% 82-90% 78-97% 79-97%

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#:Hsp27) 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. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

Multiplex Assay Kit for Heat Shock Protein 27 (Hsp27) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Anti-Heat Shock Protein 27 (Hsp27) antibody was coated on the surface of magnetic beads, to make solid phase carrier. Standard or sample were added to the magnetic beads, respectively, in which Heat Shock Protein 27 (Hsp27) was bound to the antibody connected to the solid phase carrier, and then biotinylated anti-Heat Shock Protein 27 (Hsp27) antibodies were added. PE-labeled avidin was added. There is a positive correlation between the concentration of Heat Shock Protein 27 (Hsp27) in the sample and corresponding value of MFI (Median Fluorescence Intensity).

Giveaways

Citations

  • The effect of heat stress on gene expression and synthesis of heat‐shock and milk proteins in bovine mammary epithelial cellsPubMed: 26467738
  • HSP27 is a partner of JAK2-STAT5 and a potential therapeutic target in myelofibrosisPubmed:29650953
  • Molecules of Damage-Associated Patterns in Bronchoalveolar Lavage Fluid and Serum in Chronic Obstructive Pulmonary DiseasePubmed:29429028
  • The beneficial effects of 15 units of high-intensity circuit training in women is modified by age, baseline insulin resistance and physical capacityPubmed: 31102684
  • Damage-Associated Molecular Patterns and Myeloid-Derived Suppressor Cells in Bronchoalveolar Lavage Fluid in Chronic Obstructive Pulmonary Disease Patients
  • Damage-Associated Molecular Patterns and Th-Cell-Related Cytokines Released after Progressive EffortPubmed: 32210109
  • Regulatory T cells, damage-associated molecular patterns, and myeloid-derived suppressor cells in bronchoalveolar lavage fluid interlinked with chronic obstructive …Pubmed:35687771
  • Immunization of mice with gold nanoparticles conjugated to thermostable tumor antigens prevents tumor development during transplantation

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