RPS6 ELISA Kit

Catalog No. ABIN2748617

Human and Mouse RPS6 ELISA Kit Colorimetric assay for quantification of Human and Mouse RPS6.

Quick Overview for RPS6 ELISA Kit (ABIN2748617)

Target: RPS6 (Ribosomal Protein S6 (RPS6))

Binding Specificity: pSer235, pSer236

Reactivity: Human, Mouse

Detection Method: Colorimetric

Method Type: Cell ELISA

Application: ELISA

Sample Type: Cell Culture Cells

Product Details RPS6 ELISA Kit

Purpose: Cell-Based Human/Mouse RPS6 (S235/S236) Phosphorylation ELISA Kit. Suitable for adherent whole cell lines.

Analytical Method: Semi-Quantitative

Specificity: The antibodies provided in this kit recognizes human/mouse/rat RPS6 phosphorylated at Serine-235 / Serine-236 and total RPS6 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 RPS6

Target: RPS6 (Ribosomal Protein S6 (RPS6))

Alternative Name: RPS6

Background: Disease resistance protein RPS6, Resistance to Pseudomonas syringae 6 (RPS6)

Gene ID: 6194

UniProt: P62753

Pathways: Carbohydrate Homeostasis, Ribonucleoprotein Complex Subunit Organization, Ribosome Assembly