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Manual / SDS

Description
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Antigen Information

Accession Number:

P41159

Gene ID:

3952

Gene Symbols:

Protein Name & Synonyms:

Leptin (Obese protein) (Obesity factor)

Target Species:

Human

Assay Format

Detection Range:

2 pg/ml - 400 pg/ml

Recommended Dilution (serum & plasma):

20 - 50 fold

Sensitivity:

2 pg/ml

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Specificity:

The Leptin ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, MCP- 1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.

Number of Targets Detected:

Species Detected:

Human

Compatible Sample Types:

Cell Culture Supernatants
Plasma
Serum

Design Principle:

Sandwich-based

Method of Detection:

Colorimetric

Quantitative/Semi-Quantitative:

Quantitative

Solid Support:

96-well Microplate

Product Specifications

Size:

1, 2, or 5 x 96-Well Strip Microplate Kit

Product Features

Additional Notes

Strip plates and additional reagents allow for use in multiple experiments
Quantitative protein detection
Establishes normal range
The best products for confirmation of antibody array data

Standard Curves

Spiking & Recovery Results

Linearity Results

Application Notes

Kit Components

Pre-Coated 96-well Strip Microplate
Wash Buffer
Stop Solution
Assay Diluent(s)
Lyophilized Standard
Biotinylated Detection Antibody
Streptavidin-Conjugated HRP
TMB One-Step Substrate

Other Materials Required

Distilled or deionized water
Precision pipettes to deliver 2 µl to 1 µl volumes
Adjustable 1-25 µl pipettes for reagent preparation
100 µl and 1 liter graduated cylinders
Tubes to prepare standard and sample dilutions
Absorbent paper
Microplate reader capable of measuring absorbance at 450nm
Log-log graph paper or computer and software for ELISA data analysis

Protocol Outline

Prepare all reagents, samples and standards as instructed in the manual.
Add 100 µl of standard or sample to each well.
Incubate 2.5 h at RT or O/N at 4°C.
Add 100 µl of prepared biotin antibody to each well.
Incubate 1 h at RT.
Add 100 µl of prepared Streptavidin solution to each well.
Incubate 45 min at RT.
Add 100 µl of TMB One-Step Substrate Reagent to each well.
Incubate 30 min at RT.
Add 50 µl of Stop Solution to each well.
Read at 450 nm immediately.

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Storage/Stability

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.