TRAF6 ELISA Kit

Catalog No. ABIN8071065

This Colorimetric ELISA kit is designed for the quantitative measurement of Human TRAF6.

Quick Overview for TRAF6 ELISA Kit (ABIN8071065)

Target: TRAF6 (TNF Receptor-Associated Factor 6 (TRAF6))

Binding Specificity: total, ubiquitinated

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details TRAF6 ELISA Kit

Purpose: Measures ubiquitinated TRAF6 and total TRAF6 in human cell and tissue lysates, enabling precise assessment of TRAF6 ubiquitination 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-TRAF6 Antibody
• Anti-Ubiquitination Detection Antibody
• Anti-TRAF6 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 TRAF6

Target: TRAF6 (TNF Receptor-Associated Factor 6 (TRAF6))

Alternative Name: TRAF6

Background:

Gene Symbols: TRAF6, RNF85

Background: The Ubiquitinated-TRAF6 and Total-TRAF6 ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human ubiquitinated ATP-citrate synthase and total ATP-citrate synthase. An anti-pan TRAF6 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and TRAF6 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-ubiquitination antibody is used to detect ubiquitinated TRAF6 or biotinylated anti-TRAF6 antibody is used to detect total TRAF6 . 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 ubiquitinated TRAF6 or total TRAF6 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): TNF receptor-associated factor 6, E3 ubiquitin-protein ligase TRAF6, Interleukin-1 signal transducer, RING finger protein 85, RING-type E3 ubiquitin transferase TRAF6

Gene ID: 7189

UniProt: Q9Y4K3

Pathways: NF-kappaB Signaling, TCR Signaling, TLR Signaling, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, Activation of Innate immune Response, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Tube Formation, Hepatitis C, Toll-Like Receptors Cascades, Ubiquitin Proteasome Pathway