Nori Rabbit Serpin F2/A2AP ELISA Kit

$461.00$832.00

DataSheet   

This ELISA kit is for quantification of A2AP in rabbit. This is a quick ELISA assay that reduces time to 50% compared to the conventional method, and the entire assay only takes 3 hours. This assay employs the quantitative sandwich enzyme immunoassay technique and uses biotin-streptavidin chemistry to improve the performance of the assays. An antibody specific for A2AP has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any A2AP present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for A2AP is added to the wells. Following wash to remove any unbound antibody reagent, a detection reagent is added. After intensive wash a substrate solution is added to the wells and color develops in proportion to the amount of A2AP bound in the initial step. The color development is stopped, and the intensity of the color is measured.

Alternative names for serpin F2: A2AP, alpha 2-antiplasmin, serpinf2, plasmin inhibitor

 

This product is for Laboratory Research Use Only not for diagnostic and therapeutic purposes or any other purposes.

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Description

Nori Rabbit Serpin F2/A2AP ELISA Kit Summary

Alternative names for serpin F2: A2AP, alpha 2-antiplasmin, serpinf2, plasmin inhibitor

Assay Type Solid Phase Sandwich ELISA
Format 96-well Microplate or 96-Well Strip Microplate
Method of Detection Colorimetric
Number of Targets Detected 1
Target Antiben Accession Number Q45GR2
Assay Length 3 hours
Quantitative/Semiquantitative Quantitative
Sample Type Plasma, Serum, Cell Culture, Urine, Cell/Tissue Lysates, Synovial Fluid, BAL,
Recommended Sample Dilution (Plasma/Serum) No dilution for sample <ULOQ; sufficient dilution for samples >ULOQ
Sensitivity 12 pg/mL
Detection Range 62.5-4000 pg/mL
Specificity  Rabbit A2AP
Cross-Reactivity < 0.5% cross-reactivity observed with available related molecules, < 50% cross-species reactivity observed with species tested.
Interference No significant interference observed with available related molecules
Storage/Stability 4 ºC for up to 6 months
Usage For Laboratory Research Use Only. Not for diagnostic or therapeutic use.
Additional Notes The kit allows for use in multiple experiments.

 

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Serpin F2 is also called alpha 2-antiplasmin (A2AP or plasmin inhibitor) is a serine protease inhibitor (serpin) responsible for inactivating plasmin and a member of serpin family. This protein is encoded by the SERPINF2 gene. Plasmin is an important enzyme that participates in fibrinolysis and degradation of various other proteins. Very few cases (<20) of A2AP deficiency have been described. As plasmin degrades blood clots, impaired inhibition of plasmin leads to a bleeding tendency, which was severe in the cases reported. In liver cirrhosis, there is decreased production of alpha 2-antiplasmin, leading to decreased inactivation of plasmin and an increase in fibrinolysis. This is associated with an increased risk of bleeding in liver disease.[1] Alpha 2-antiplasmin has been shown to interact with neutrophil elastase[2][3] and plasmin.[2][4]

References

  1. Sattar, Husain. Fundamentals of Pathology. Pathoma LLC, 2011, p. 36.
  2. Shieh BH, Travis J (May 1987). “The reactive site of human alpha 2-antiplasmin”. The Journal of Biological Chemistry. 262 (13): 6055–9.
  3. Brower MS, Harpel PC (Aug 1982). “Proteolytic cleavage and inactivation of alpha 2-plasmin inhibitor and C1 inactivator by human polymorphonuclear leukocyte elastase”. The Journal of Biological Chemistry. 257 (16): 9849–54.
  4. Wiman B, Collen D (Sep 1979). “On the mechanism of the reaction between human alpha 2-antiplasmin and plasmin”. The Journal of Biological Chemistry. 254 (18): 9291–7.

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