HIF1A ELISA Kit

Catalog No. ABIN5690734

This Colorimetric ELISA kit is designed for the quantitative measurement of Human HIF1A.

Quick Overview for HIF1A ELISA Kit (ABIN5690734)

Target: HIF1A (Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) (HIF1A))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Cell ELISA

Application: ELISA

Sample Type: Adherent Cell Culture

Product Details HIF1A ELISA Kit

Purpose: 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.

Analytical Method: Semi-Quantitative

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

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

Plate: Uncoated

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: 4 °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 HIF1A

Target: HIF1A (Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) (HIF1A))

Alternative Name: HIF-1alpha

Gene ID: 3091

UniProt: Q16665

Pathways: Positive Regulation of Peptide Hormone Secretion, Regulation of Hormone Metabolic Process, Regulation of Hormone Biosynthetic Process, Cellular Response to Molecule of Bacterial Origin, Carbohydrate Homeostasis, Transition Metal Ion Homeostasis, Tube Formation, Regulation of Carbohydrate Metabolic Process, Signaling Events mediated by VEGFR1 and VEGFR2, VEGFR1 Specific Signals, Warburg Effect