TLR4 ELISA Kit

Catalog No. ABIN8071080

This Colorimetric ELISA kit is designed for the quantitative measurement of Human TLR4.

Quick Overview for TLR4 ELISA Kit (ABIN8071080)

Target: TLR4 (Toll-Like Receptor 4 (TLR4))

Binding Specificity: nitrated, total

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details TLR4 ELISA Kit

Purpose: Measures nitrated TLR4 and total TLR-4 in human cell and tissue lysates, enabling precise assessment of TLR-4 nitration by ELISA. This assay can be developed for other species (mouse, rat, etc.) upon request. Contact techsupport@raybiotech.com.

Analytical Method: Quantitative

Components:

Size / Qty
• Microplate Coated With Anti-Pan-TLR-4 Antibody
• Anti-Nitration Detection Antibody
• Anti-TLR-4 Detection Antibody
• HRP-Conjugated Streptavidin
• 200 μL
• Wash Buffer
• Assay Diluent
• 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 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 and samples as instructed.
2. Add 100 μL 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-Conjugated 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.

Target Details for TLR4

Target: TLR4 (Toll-Like Receptor 4 (TLR4))

Alternative Name: TLR4

Background:

Gene Symbols: TLR4

Background: The Nitrated-TLR-4 and Total-TLR-4 ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human nitrated ATP-citrate synthase and total ATP-citrate synthase. An anti-pan TLR-4 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and TLR-4 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-nitration antibody is used to detect nitrated TLR-4 or biotinylated anti-TLR-4 antibody is used to detect total TLR-4 . After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to 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 nitrated TLR-4 or total TLR-4 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): Toll-like receptor 4, hToll

Gene ID: 7099

UniProt: O00206

Pathways: TLR Signaling, Activation of Innate immune Response, Cellular Response to Molecule of Bacterial Origin, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Toll-Like Receptors Cascades, Inflammasome, S100 Proteins