Nori Human Apelin 12 ELISA Kit

$508.00$916.00

DataSheet   

This ELISA kit is for quantification of apelin 12 in human. 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 APLN12 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any APLN12 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for APLN12 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 APLN12 bound in the initial step. The color development is stopped, and the intensity of the color is measured.

Alternative names for apelin 12: APLN12, APLN 12

This product is for laboratory research use only not for diagnostic and therapeutic purposes or any other purposes.

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Description

Nori Human Apelin 12 ELISA Kit Summary

Alternative names for apelin 12: APLN 12, APLN12

 

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 Antigen Accession Number NA
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 6 pg/mL
Detection Range 31.25-2000 pg/mL
Specificity Human apelin 12
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:

Apelin (also known as APLN) is a peptide that in humans is encoded by the APLN gene.[1] Apelin is the endogenous ligand for the G-protein-coupled APJ receptor[2-6] that is expressed at the surface of some cell types. It is widely expressed in various organs such as the heart, lung, kidney, liver, adipose tissue, gastrointestinal tract, brain, adrenal glands, endothelium, and human plasma. Apelin is a new peptide that was identified in 1998 by Professor M. Fujino’s team.[1] Apelin gene encodes a pre-proprotein of 77 amino acids,[1] with a signal peptide in the N-terminal region. After translocation into the endoplasmic reticulum and cleavage of the signal peptide, the proprotein of 55 amino acids may generate several active fragments: a 36 amino acid peptide corresponding to the sequence 42-77 (apelin 36), a 17 amino acid peptide corresponding to the sequence 61-77 (apelin 17) and a 13 amino acid peptide corresponding to the sequence 65-77 (apelin 13). This latter fragment may also undergo a pyroglutamylation at the level of its N-terminal glutamine residue. However the presence and/or the concentrations of those peptides in human plasma has been questioned.[7] Recently, 46 different apelin peptides ranging from apelin 55 (proapelin) to apelin 12 have been identified in bovine colostrum, including C-ter truncated isoforms.[8] The sites of receptor expression are clearly linked to the different functions played by apelin in the organism.

References

  1. Tatemoto K, et al. (1998). Biophys. Res. Commun. 251 (2): 471–6.
  2. Lee DK, et al. (2000). Neurochem. 74 (1): 34–41.
  3. Szokodi I, et al. (2002). Res. 91 (5): 434–40.
  4. Kleinz MJ, et al. (2005). Ther. 107 (2): 198–211.
  5. O’Dowd BF, et al. (1993). Gene 136 (1–2): 355–60.
  6. Devic E, et al. (1996). Dev. 59 (2): 129–40.
  7. Mesmin C, et al. (2010). Rapid Commun Mass Spectrom 24 (19): 2875–84.
  8. Mesmin C, et al. (2011). J Proteome Res 10 (11): 5222–31.

 

 

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