p53 ELISA Kit

Catalog No. ABIN2748488

Human, Rat and Mouse p53 ELISA Kit Colorimetric assay for quantification of Human, Rat and Mouse p53 and has been mentioned in 1 publication.

Quick Overview for p53 ELISA Kit (ABIN2748488)

Target: p53 (TP53) (Tumor Protein P53 (TP53))

Binding Specificity: pSer15

Reactivity: Human, Rat, Mouse

Detection Method: Colorimetric

Method Type: Cell ELISA

Application: ELISA

Sample Type: Cell Culture Cells

Product Details p53 ELISA Kit

Purpose: Cell-Based Human/Rat/Mouse p53 (S15) Phosphorylation ELISA Kit. Suitable for adherent whole cell lines.

Analytical Method: Semi-Quantitative

Specificity: The antibodies provided in this kit recognizes Human/Rat/Mouse p53 phosphorylated at Serine-15 and total p53 for comparison.

Characteristics:

• Site and signal pathway-specific
• In vitro detection of adherent cell culture
• No sample lysis needed
• Compatible with a standard ELISA plate reader
• Faster results than with ELISA
• Adaptable for high-throughput screening and drug discovery

Components:

• uncoated 96-well Microplate
• Wash Buffer A
• Wash Buffer B
• Fixing Solution
• Quenching Buffer
• Blocking Buffer
• Anti-phospho antibody
• Anti-pan antibody
• HRP-Conjugated Secondary Antibody
• TMB One-Step Substrate
• Stop Solution

Material not included:

• Distilled or deionized water
• 100 mL and 1 liter graduated cylinders
• Tubes to prepare sample dilutions
• Protease and Phosphatase inhibitors
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• Benchtop rocker or shaker
• Microplate reader capable of measuring absorbance at 450 nm

Application Details

Sample Volume: 100 μL

Plate: Uncoated

Protocol:

1. Seed 10,000-30,000 cells into each well and incubate overnight.
2. Apply various treatment, inhibitors or activators according to manufacture's instructions.
3. Add 100 μL of Fixing Solution into each well and incubate for 20 min at RT with shaking.
4. Add 200 μL of prepared 1X Quenching Buffer and incubate 20 min at RT.
5. Add 200 μL of Blocking Solution and incubate for 1 h at 37 °C.
6. Add 50 μL of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.
7. Add 50 μL of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Assay Procedure:

Seed 10,000-30,000 cells into each well and incubate overnight.Apply various treatment, inhibitors or activators according to manufacture's instructions.Add 100 μL of Fixing Solution into each well and incubate for 20 min at RT with shaking.Add 200 μL of prepared 1X Quenching Buffer and incubate 20 min at RT.Add 200 μL of Blocking Solution and incubate for 1 h at 37 °C.Add 50 μL of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.Add 50 μL of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.Add 100 μL of TMB One-Step Substrate Reagent to each well.Incubate 30 min at RT.Add 50 μL of 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.

Expiry Date: 6 months

References for p53 ELISA Kit (ABIN2748488)

Hassan, Khan, Aman, Qamar, Ebaid, Al-Tamimi, Alhazza, Rady: "Restrained management of copper level enhances the antineoplastic activity of imatinib in vitro and in vivo." in: Scientific reports, Vol. 8, Issue 1, pp. 1682, (2018) (PubMed).

Target Details for p53

Target: p53 (TP53) (Tumor Protein P53 (TP53))

Alternative Name: P53

Background: Cellular tumor antigen p53, Antigen NY-CO-13, Phosphoprotein p53, Tumor suppressor p53

Gene ID: 7157

UniProt: P04637

Pathways: p53 Signaling, MAPK Signaling, PI3K-Akt Signaling, Apoptosis, AMPK Signaling, Chromatin Binding, ER-Nucleus Signaling, Positive Regulation of Endopeptidase Activity, Hepatitis C, Protein targeting to Nucleus, Autophagy, Warburg Effect