Custom Human Hydroxylated-HIF-1alpha (P402) Cell-based ELISA

RayBio® Human Hydroxylated-HIF-1 alpha (P402) Cell-based ELISA Kit. This ELISA is for measuring hydroxylated HIF-1 alpha (Pro-402) in human adherent cell lines.
Lead time 4-6 weeks

$633.00  
CBEL-HIF1A-P402-1
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Antigen Information

Accession Number:
  • Q16665
Gene ID:
  • 3091
Gene Symbols:
  • BHLHE78
  • HIF1A
  • MOP1
  • PASD8
Protein Name & Synonyms:
Hypoxia-inducible factor 1-alpha (HIF-1-alpha) (HIF1-alpha) (ARNT-interacting protein) (Basic-helix-loop-helix-PAS protein MOP1) (Class E basic helix-loop-helix protein 78) (bHLHe78) (Member of PAS protein 1) (PAS domain-containing protein 8)
Target Species:
  • Human

Assay Format

Specificity:
The antibody pair provided in this kit recognizes human Hydroxylated-HIF-1alpha hydroxylated at site Proline-402

Product Specifications

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

Product Features

  • Rapidly measure Hydroxylated protein in lysates
  • Simultaneously measure hydroxylated 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
  • Quneching Buffer
  • Blocking Buffer
  • Anti-cleaved 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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Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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