PCNA ELISA Kit

Catalog No. ABIN8070506

Human PCNA ELISA Kit, Colorimetric assay for quantification of Human PCNA.

Quick Overview for PCNA ELISA Kit (ABIN8070506)

Target: PCNA (Proliferating Cell Nuclear Antigen (PCNA))

Binding Specificity: SUMOylated, total

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details PCNA ELISA Kit

Purpose: Measures sumoylated PCNA and total PCNA in human cell and tissue lysates, enabling precise assessment of PCNA 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-PCNA Antibody
• Anti-SUMOylation Detection Antibody
• Anti-PCNA 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 PCNA

Target: PCNA (Proliferating Cell Nuclear Antigen (PCNA))

Alternative Name: PCNA

Background:

Gene Symbols: PCNA

Background: The SUMOylated-PCNA and Total-PCNA 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 PCNA antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and PCNA 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 PCNA or biotinylated anti-PCNA antibody is used to detect total PCNA . 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 PCNA or total PCNA 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): Proliferating cell nuclear antigen (PCNA), Cyclin

Gene ID: 5111

UniProt: P12004

Pathways: Telomere Maintenance, DNA Damage Repair, Mitotic G1-G1/S Phases, DNA Replication, Synthesis of DNA, Autophagy