MAP2K3 ELISA Kit

Catalog No. ABIN2748359

Human, Mouse and Rat MAP2K3 ELISA Kit Colorimetric assay for quantification of Human, Mouse and Rat MAP2K3.

Quick Overview for MAP2K3 ELISA Kit (ABIN2748359)

Target: MAP2K3 (Mitogen-Activated Protein Kinase Kinase 3 (MAP2K3))

Binding Specificity: pSer189, total

Reactivity: Human, Mouse, Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details MAP2K3 ELISA Kit

Purpose: Human/Mouse/Rat Phospho-MKK3 (S189) and Total MKK3 ELISA Kit. This assay semi-quantitatively measures phophorylated MKK3 (Ser189) and Total MKK3 in lysate samples.

Analytical Method: Semi-Quantitative

Specificity: The antibody pair provided in this kit recognizes human, mouse, and rat MKK3 phosphorylated at site Serine-189 and Total MKK3.

Characteristics:

• Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
• Screen numerous different cell lysates without performing a Western Blot analysis
• Minimal hands-on time, convenient, and non-radioactive material

Components:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Anti-Phospho Antibody
• Anti-Pan Antibody
• HRP-Conjugated Secondary Antibody
• Streptavidin-Conjugated HRP
• Assay Diluent
• TMB One-Step Substrate
• Stop Solution
• Lysis Buffer
• Positive Control Sample

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: Pre-coated

Protocol:

1. Prepare all reagents and samples as instructed in the manual.
2. Add 100 μL of sample or positive control to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared primary antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared 1X HRP-Streptavidin to each well.
7. Incubate 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:

Prepare all reagents and samples as instructed in the manual.
Add 100 μL of sample or positive control to each well.
Incubate 2.5 h at RT or O/N at 4 °C.
Add 100 μL of prepared primary antibody to each well.
Incubate 1 h at RT.
Add 100 μL of prepared 1X HRP-Streptavidin to each well.
Incubate 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: Upon receipt, the kit should be stored at -20 °C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), HRP-Streptavidin (Item G), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4 °C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge and store at -20 °C. Reconstituted Positive Control (Item K) should be stored at -70 °C.

Expiry Date: 6 months

Target Details for MAP2K3

Target: MAP2K3 (Mitogen-Activated Protein Kinase Kinase 3 (MAP2K3))

Alternative Name: MKK3

Background: Mitogen-activated protein kinase kinase 3 (MKK3) phosphorylated at Serine-189 and Total MKK3

Gene ID: 5606

UniProt: P46734

Pathways: MAPK Signaling, TLR Signaling, Activation of Innate immune Response, Toll-Like Receptors Cascades, Autophagy, Signaling Events mediated by VEGFR1 and VEGFR2