HIF1A ELISA Kit

Catalog No. ABIN4889776

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

Quick Overview for HIF1A ELISA Kit (ABIN4889776)

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

Reactivity: Human

Detection Method: Colorimetric

Method Type: DNA-Binding ELISA

Application: ELISA

Sample Type: Cell Lysate, Nuclear Extract

Product Details HIF1A ELISA Kit

Purpose: Human HIF-1α Transcription Factor Activity Assay. This assay uses a dsDNA coated plate with canonical HIF-1α binding sequences to semi-quantitatively detect active HIF-1α in lysates or nuclear extracts. Only available in North America.

Analytical Method: Semi-Quantitative

Specificity: The olionucleotide/antibody pair provided in this kit recognizes human HIF-1α in whole lysates and nuclear extracts.

Characteristics:

• Specific transcription factor-DNA binding assay
• Perfect alternative to EMSA
• Easy to perform in an ELISA format
• Non-radioactive assay
• High throughput (96 well plate format)
• Assay can be completed within 5 hours

Components:

• 96-well Strip Microplate pre-coated with DNA probes
• DNA Binding Buffer
• Positive Control Sample
• Specific Competitor DNA probe
• Non-specific Competitor DNA probe
• Assay Reagent
• DTT
• Wash Buffer
• Primary Antibody
• HRP-conjugated Secondary Antibody
• TMB One-Step Substrate Reagent
• Stop Solution

Material not included:

• Distilled or deionized water
• 100 mL and 1 liter graduated cylinders
• Tubes to prepare sample dilutions Absorbent paper
• 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.

Sample Volume: 100 μL

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 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 HRP-secondary antibody 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 positive control should be removed and stored at -20° or -80°C. The remainder of the kit can be stored for up to 6 months at 2-8°C from the date of shipment. Opened Microplate Wells or reagents may be stored for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack, reseal along entire edge.
Note: The kit can be used within one year if the whole kit is stored at -20°C upon receipt. Avoid repeated freeze-thaw cycles.

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-1 alpha

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