PTPN11 ELISA Kit

Catalog No. ABIN2747888

Human and Mouse PTPN11 ELISA Kit, Colorimetric assay for quantification of Human and Mouse PTPN11.

Quick Overview for PTPN11 ELISA Kit (ABIN2747888)

Target: PTPN11 (Protein tyrosine Phosphatase, Non-Receptor Type 11 (PTPN11))

Binding Specificity: pTyr542

Reactivity: Human, Mouse

Detection Method: Colorimetric

Method Type: Cell ELISA

Application: ELISA

Sample Type: Cell Culture Cells

Product Details PTPN11 ELISA Kit

Purpose: Cell-Based Human/Mouse SHP2 (Y542) Phosphorylation ELISA Kit. Suitable for adherent whole cell lines.

Analytical Method: Semi-Quantitative

Specificity: The antibodies provided in this kit recognizes human and mouse SHP2 phosphorylated at Tyrosine-542 and total SHP2 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

Target Details for PTPN11

Target: PTPN11 (Protein tyrosine Phosphatase, Non-Receptor Type 11 (PTPN11))

Alternative Name: SHP2

Target Type: Viral Protein

Background: Tyrosine-protein phosphatase non-receptor type 11, Protein-tyrosine phosphatase 1D (PTP-1D), Protein-tyrosine phosphatase 2C (PTP-2C), SH-PTP2 (SHP-2), SH-PTP3

Gene ID: 5781

UniProt: Q06124

Pathways: JAK-STAT Signaling, RTK Signaling, TCR Signaling, Interferon-gamma Pathway, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Negative Regulation of Hormone Secretion, Carbohydrate Homeostasis, Toll-Like Receptors Cascades, CXCR4-mediated Signaling Events, Signaling Events mediated by VEGFR1 and VEGFR2, Signaling of Hepatocyte Growth Factor Receptor, VEGFR1 Specific Signals, BCR Signaling, Warburg Effect