Human Glycocalicin ELISA kit | CSB-E12101h

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SKU:
CSB-E12101h
Availability:
3-5 working days
Species:
Homo sapiens (Human)
Sample Types:
serum, plasma, tissue homogenates
Detection Range:
0.156 μg/mL-10 μg/mL
Sensitivity:
0.039 μg/mL
Assay Time:
1-5h
Sample Volume:
50-100ul
Detection Wavelength:
450 nm
Research Area:
Blood Coagulation
Assay Principle:
quantitative
Measurement:
Sandwich
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Description

CUSABIO’s Human Glycocalicin ELISA kit is designed for the quantitative detection of human glycocalicin in serum, plasma, or tissue homogenates. This kit has been validated with high sensitivity, excellent specificity, good linearity, high recovery, precision low than 10%, and high lot-to-lot consistency. The detection mechanism of this kit is based on the sandwich ELISA technique and enzyme-substrate chromogenic reaction. The solution color develops in proportion to the amount of glycocalicin in the sample, and the intensity of the color can be measured at 450 nm via a microplate reader.

Platelet glycocalicin is the major and extramembranous domain of Glycoprotein Iba (GPlba) that can be rapidly cleaved by proteases such as calpain, plasmin, and trypsin. It is not only a marker but also a parameter that reflects platelet turnover, therefore being an aid in the classification of thrombocytopenic disorders. Besides, platelet glycocalicin could act as a circulating inhibitor of thrombin. The levels of platelet glycocalicin were elevated in patients with myocardial infarction after receiving recombinant tissue plasminogen activator (rtPA) treatment. Platelet glycocalicin levels in platelet concentrates may be helpful to qualitative control the storage conditions over time for blood banks.

Target Name: glycoprotein Ib (platelet), alpha polypeptide

Alternative Names: GP1BA ELISA Kit; Platelet glycoprotein Ib alpha chain ELISA Kit; GP-Ib alpha ELISA Kit; GPIb-alpha ELISA Kit; GPIbA ELISA Kit; Glycoprotein Ibalpha ELISA Kit; Antigen CD42b-alpha ELISA Kit; CD antigen CD42b) [Cleaved into: Glycocalicin] ELISA Kit

Abbreviation:  GP1BA

Uniprot No.:    P07359

Species:    Homo sapiens (Human)

Sample Types:  serum, plasma, tissue homogenates

Detection Range:  0.156 μg/mL-10 μg/mL

Sensitivity:  0.039 μg/mL

Assay Time: 1-5h

Sample Volume:  50-100ul

Detection Wavelength:   450 nm

Research Area:    Blood Coagulation

Assay Principle:  quantitative

Measurement:  Sandwich

Manual:    Download PDF Manual

Precision: 

Intra-assay Precision (Precision within an assay): CV%<8%      
Three samples of known concentration were tested twenty times on one plate to assess.  
Inter-assay Precision (Precision between assays): CV%<10%      
Three samples of known concentration were tested in twenty assays to assess.    
   

Linearity:

To assess the linearity of the assay, samples were spiked with high concentrations of human glycocalicin in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
  Sample Serum(n=4)  
1:1 Average % 90  
Range % 87-94  
1:2 Average % 95  
Range % 91-99  
1:4 Average % 84  
Range % 80-88  
1:8 Average % 86  
Range % 82-90

Recovery:

The recovery of human glycocalicin spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample Type Average % Recovery Range  
Serum (n=5) 89 84-93  
EDTA plasma (n=4) 92 87-96

Typical Data: These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.

μg/ml. OD1 OD2 Average Corrected  
10 2.246 2.268 2.257 2.130  
5 1.395 1.374 1.385 1.258  
2.5 0.766 0.798 0.782 0.655  
1.25 0.488 0.462 0.475 0.348  
0.625 0.326 0.354 0.340 0.213  
0.312 0.220 0.233 0.227 0.100  
0.156 0.168 0.164 0.166 0.039  
0 0.126 0.128 0.127  

Materials provided:

  • A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-human glycocalicin. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled glycocalicin antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual

Materials not provided:

  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution

Storage: Store at 2-8°C. Please refer to protocol.

Lead Time: 3-5 working days

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