SIRT1 ELISA Kit

Catalog No. ABIN8070804

This Colorimetric ELISA kit is designed for the quantitative measurement of Human SIRT1.

Quick Overview for SIRT1 ELISA Kit (ABIN8070804)

Target: SIRT1 (Sirtuin 1 (SIRT1))

Binding Specificity: SUMOylated, total

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details SIRT1 ELISA Kit

Purpose: Measures sumoylated SIRT1 (Sirtuin 1) and total SIRT1 in human cell and tissue lysates, enabling precise assessment of SIRT1 sumoylation 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-SIRT1 Antibody
• Anti-SUMOylation Detection Antibody
• Anti-SIRT1 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 SIRT1

Target: SIRT1 (Sirtuin 1 (SIRT1))

Alternative Name: Sirtuin 1

Background:

Gene Symbols: SIRT1, SIR2L1

Background: The SUMOylated-SIRT1 and Total-SIRT1 ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human sumoylated ATP-citrate synthase and total ATP-citrate synthase. An anti-pan SIRT1 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and SIRT1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-sumoylation antibody is used to detect sumoylated SIRT1 or biotinylated anti-SIRT1 antibody is used to detect total SIRT1 . 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 sumoylated SIRT1 or total SIRT1 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): NAD-dependent protein deacetylase sirtuin-1 (hSIRT1), NAD-dependent protein deacylase sirtuin-1, Regulatory protein SIR2 homolog 1, SIR2-like protein 1 (hSIR2)

Gene ID: 23411

UniProt: Q96EB6

Pathways: MAPK Signaling, Intracellular Steroid Hormone Receptor Signaling Pathway, Regulation of Intracellular Steroid Hormone Receptor Signaling, Carbohydrate Homeostasis, Positive Regulation of Endopeptidase Activity, Regulation of Carbohydrate Metabolic Process, Positive Regulation of Response to DNA Damage Stimulus, Negative Regulation of intrinsic apoptotic Signaling