HIF1A IQ-ELISA Kit

Catalog No. ABIN6385981

Product Details HIF1A IQ-ELISA Kit

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

Reactivity: Human, Mouse

Detection Method: qPCR

Method Type: Sandwich ELISA

Application: ELISA

Purpose: Human Immunoquantitative (PCR-Based) HIF-1 alpha ELISA Kit for cell culture supernatants, plasma, and serum samples.

Sample Type: Cell Culture Supernatant, Plasma, Serum

Analytical Method: Semi-Quantitative

Sensitivity: 6.1 pg/mL

Characteristics: IQ-ELISAs employ specific capture antibodies coated on a 96-well PCR plate. Standards and samples are pipetted into the wells, the target protein in the standards and samples binds to the immobilized antibody. The wells are washed and the detection affinity reagent is added to the wells where it binds to any captured antigen. The wells are washed, and primers and PCR master mix are added to each well. The plate is placed into a real time PCR instrument for cycling and measurement of DNA amplification. The cycle number where amplification is detected is proportional to the amount of affinity detection reagent that bound to captured antigen in each well.

Components:

• HIF-1 alpha Microplate (Item A): 96 well PCR plate coated with anti-Human HIF-1 alpha
• Wash Buffer I Concentrate (20x) (Item B): 25 ml of 20x concentrated solution
• Standards (Item C): 2 vials of recombinant Human HIF-1 alpha
• Assay Diluent A (Item D): 30 ml diluent buffer, 0.09% sodium azide as preservative. For Standard/Sample (serum/plasma) diluent
• Assay Diluent B (Item E): 15 ml of 5x concentrated buffer. For Standard/Sample (cell culture medium/urine) diluent
• Detection Affinity Reagent for HIF-1 alpha (Item F): 2 vials of a 4x concentrated solution of anti-Human HIF-1 alpha affinity reagent
• IQELISA Detection Reagent (Item G): 1.4ml of a 10x concentrated stock
• Primer Solution (Item I): 1.7 ml vial
• PCR Master Mix (Item J): 1.2 ml vial
• PCR Preparation buffer (Item K): 1ml vial of 10x concentrated buffer
• Final Wash Buffer (Item L): 10ml vial of 10x concentrated buffer

Material not included:

• Real-time PCR instrument, Bio-Rad recommended
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• 100 mL and 1 liter graduated cylinders
• Absorbent paper
• Distilled or deionized water
• Log-log graph paper or computer and software for data analysis
• Tubes to prepare standard or sample dilutions
• Heating block or water bath capable of 80^°C

Application Details

Application Notes: The Immuno-Quantitative ELISA (IQELISA) kits are an innovative assay platform that combines the specificity and ease of use of an ELISA with the sensitivity of real-time PCR. This results in an assay that is simultaneously familiar and cutting edge and enables the use of only 1/10th the sample volume while also providing 10x more sensitivity than a traditional ELISA.

Comment:

The Immuno-Quantitative ELISA (IQELISA™) kits are an innovative assay platform that combines the specificity and ease of use of an ELISA with the sensitivity of real-time PCR. Also called immuno-PCR, this detection platform results in an assay that is simultaneously familiar and cutting edge. Compared to traditional ELISA, IQELISA™ enables the use of only 1/10th the sample volume while also providing 10x more sensitivity.

Sample Volume: 25 µL

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and standards as instructed
2. Add 25 μL standard or sample to each well. Incubate for 2.5 hours at room temperature or overnight at 4^°C
3. Add 25 μL detection affinity reagent to each well. Incubate 1 hour at room temperature
4. Add 25μL of IQELISA Detection Reagent to each well. Incubate 1 hour
5. Add 15μL Primer solution 10μL of PCR master mix to each well
6. Run real-time PCR

Restrictions: For Research Use only

Handling

Storage: 4 °C,-20 °C,-80 °C

Storage Comment: May be stored for up to 6 months at 2° to 8°C from the date of shipment. Standard (recombinant protein) should be stored at -20°C or -80°C (recommended at -80°C) after reconstitution. Opened PCR plate or reagents may be stored for up to 1 month at 2° to 8°C. Note: the kit can be used within one year if the whole kit is stored at -20°C. Avoid repeated freeze-thaw cycles.

Expiry Date: 6 months

Target Details

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

Alternative Name: HIF-1 alpha (HIF1A Products)

Synonyms: hif1a IQ-ELISA Kit, AA959795 IQ-ELISA Kit, HIF1alpha IQ-ELISA Kit, MOP1 IQ-ELISA Kit, bHLHe78 IQ-ELISA Kit, HIF-1A IQ-ELISA Kit, HIF-1alpha IQ-ELISA Kit, HIF1 IQ-ELISA Kit, HIF1-ALPHA IQ-ELISA Kit, PASD8 IQ-ELISA Kit, HIF1-alpha IQ-ELISA Kit, hif-1a IQ-ELISA Kit, hypoxia inducible factor 1 alpha IQ-ELISA Kit, hypoxia inducible factor 1, alpha subunit IQ-ELISA Kit, hypoxia inducible factor 1 alpha subunit IQ-ELISA Kit, HIF (Hypoxia Inducible Factor) homologa IQ-ELISA Kit, hif-1a IQ-ELISA Kit, Hif1a IQ-ELISA Kit, HIF1A IQ-ELISA Kit, hif1a IQ-ELISA Kit, hif-1 IQ-ELISA Kit

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