NEFH ELISA Kit

Catalog No. ABIN8070229

The Human NEFH ELISA Kit (ABIN8070229) is a Colorimetric ELISA Kit designed to quantify Human NEFH.

Quick Overview for NEFH ELISA Kit (ABIN8070229)

Target: NEFH (Neurofilament, Heavy Polypeptide (NEFH))

Binding Specificity: phosphorylated

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.2 pg/mL - 40 ng/mL

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details NEFH ELISA Kit

Minimum Detection Limit: 0.2 pg/mL

Purpose: Phospho-NFH Quantitative ELISA kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human phosphorylated neurofilament heavy polypeptide (NFH) in lysate samples.

Analytical Method: Quantitative

Specificity: This ELISA antibody pair detects human Phospho NFH.

Components:

• Anti-Pan-NFH Microplate
• Phospho-NFH Standard
• Anti-Phospho NFH Detection Antibody
• HRP-Streptavidin
• Wash Buffer
• Assay Diluent B
• Lysis Buffer
• TMB One-Step Substrate Reagent
• Stop Solution

,

• Anti-Pan-NFH Microplate
• Phospho-NFH Standard
• Anti-Phospho NFH Detection Antibody
• HRP-Streptavidin
• Wash Buffer
• Assay Diluent B
• Lysis Buffer
• TMB One-Step Substrate Reagent
• Stop Solution

Material not included:

• Microplate reader capable of measuring absorbance at 450 nm
• Protease and Phosphatase inhibitors
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• 100 mL and 1 liter graduated cylinders
• Log-log graph paper or computer and software for ELISA data analysis
• Absorbent paper
• Tubes to prepare the positive control or sample dilutions
• Distilled or deionized water
• Shaker

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and positive control as instructed.
2. Add 100 μL of each standard and sample to each well. Incubate 2.5 hours at room temperature or overnight at 4 °C with gentle shaking.
3. Add 100 μL prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
4. Add 100 μL prepared HRP-Streptavidin solution. Incubate for 1 hour at room temperature with gentle shaking.
5. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
6. Add 50 μL Stop Solution to each well. Read at 450 nm immediately.

Restrictions: For Research Use only

Handling

Storage: -20 °C

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

Target Details for NEFH

Target: NEFH (Neurofilament, Heavy Polypeptide (NEFH))

Alternative Name: NFH

Background:

Gene Symbols: NEFH, KIAA0845, NFH

Background: Phospho-NFH Quantitative ELISA kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human phosphorylated neurofilament heavy polypeptide (NFH). An anti-pan NFH antibody has been coated onto a 96-well plate. Standard and Samples are pipetted into the wells and NFH present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-phospho-NFH (KSP site) antibody is used to detect phosphorylated NFH. After washing away unbound antibody, HRP-conjugated streptavidin is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of NFH bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Protein Name (Synonyms): Neurofilament heavy polypeptide (NF-H) (200 kDa neurofilament protein) (Neurofilament triplet H protein)

Gene ID: 4744

UniProt: P12036