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RayBiotech/Human Phospho-Caveolin-1 (Tyr14) and Total Caveolin-1 Cell-Based ELISA/1 Plate Kit/CBEL-Caveolin1-Y14-1

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面议
货号:CBEL-Caveolin1-Y14-1
浏览量:127
品牌:RayBiotech
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商品描述

Antigen Information

Accession Number:
  • Q03135
Gene ID:
  • 857
Gene Symbols:
  • CAV
  • CAV1
Protein Name & Synonyms:
Caveolin-1
Target Species:
  • Human

Assay Format

Specificity:
This ELISA kit recognizes Human Caveolin-1 phosphorylated at site Tyrosine-14 as well as total Caveolin-1.
Number of Targets Detected:
2
Species Detected:
  • Human
Compatible Sample Types:
  • Adherent Cell Culture
Design Principle:
  • Cell-based
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
  • Semi-Quantitative
Solid Support:
96-well Microplate

Product Specifications

Size:
1, 2, or 5 x 96-Well Strip Microplate Kit

Product Features

  • 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

Application Notes

Kit 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
Other Materials Required
  • 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
Protocol Outline
  1. Seed 10,000-30,000 cells into each well and incubate overnight.
  2. Apply various treatment, inhibitors or activators according to manufacturer"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.

Storage/Stability

Upon receipt, the kit should be stored at –20°C. Please use within 6 months from the date of shipment.

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