PDPK1 ELISA Kit

Catalog No. ABIN5526686

Mouse, Human and Rat PDPK1 ELISA Kit, Colorimetric assay for quantification of Mouse, Human and Rat PDPK1.

Quick Overview for PDPK1 ELISA Kit (ABIN5526686)

Target: PDPK1 (3-phosphoinositide Dependent Protein Kinase-1 (PDPK1))

Binding Specificity: pSer241, total

Reactivity: Mouse, Human, Rat

Detection Method: Colorimetric

Method Type: Cell ELISA

Application: ELISA

Sample Type: Adherent Cell Culture

Product Details PDPK1 ELISA Kit

Purpose: Cell-Based ELISA Kit. This assay semi-quantitatively measures PDK1 phosphorylated at Serine-241 as well as total PDK1 in adherent cell lines.

Analytical Method: Semi-Quantitative

Specificity: This ELISA kit recognizes Human, Mouse and Rat PDK1 phosphorylated at site Serine-241 as well as total PDK1.

Characteristics:

• Rapidly measure phosphorylated protein in adherent cell lines
• Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
• No sample lysis is needed
• Compatible with a standard ELISA plate reader

Components:

• Uncoated 96-well Strip Microplate
• Wash Buffers
• 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

Application Notes: Optimal working dilution should be determined by the investigator.

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.

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 PDPK1

Target: PDPK1 (3-phosphoinositide Dependent Protein Kinase-1 (PDPK1))

Alternative Name: PDK1

Gene ID: 5170

UniProt: O15530, Q9Z2A0, O55173

Pathways: PI3K-Akt Signaling, TCR Signaling, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Cell-Cell Junction Organization, Regulation of Cell Size, Skeletal Muscle Fiber Development, CXCR4-mediated Signaling Events, Signaling Events mediated by VEGFR1 and VEGFR2, VEGFR1 Specific Signals