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AB326404

Human HIF-2-alpha ELISA Kit, Chemiluminescent

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Human HIF-2-alpha ELISA Kit, Chemiluminescent is a 55-min single-wash SimpleStep Ignite™ ELISA for the rapid quantification of Human HIF-2-alpha with higher sensitivity and broader measuring range.

- Single 30 min incubation and single-wash step for results in less than an hour.
- Chemiluminescent detection allows a broad dynamic range: 1.83 - 4000 pg/ml with a sensitivity of 1.32pg/mL.
- Design your own immunoassay: we also offer the conjugation-ready antibody pair.
-Patent pending

View Alternative Names

BHLHE73, HIF2A, MOP2, PASD2, EPAS1, Endothelial PAS domain-containing protein 1, EPAS-1, Basic-helix-loop-helix-PAS protein MOP2, Class E basic helix-loop-helix protein 73, HIF-1-alpha-like factor, Hypoxia-inducible factor 2-alpha, Member of PAS protein 2, PAS domain-containing protein 2, bHLHe73, HLF, HIF-2-alpha, HIF2-alpha

5 Images
Sandwich ELISA - Human HIF-2-alpha ELISA Kit, Chemiluminescent (AB326404)
  • sELISA

Lab

Sandwich ELISA - Human HIF-2-alpha ELISA Kit, Chemiluminescent (AB326404)

SimpleStep ELISA® Ignite technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 55 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

Sandwich ELISA - Human HIF-2-alpha ELISA Kit, Chemiluminescent (AB326404)
  • sELISA

Lab

Sandwich ELISA - Human HIF-2-alpha ELISA Kit, Chemiluminescent (AB326404)

Dynamic range comparison between the Chemiluminescent Human HIF-2-alpha SimpleStep ELISA, colorimetric Human HIF-2-alpha SimpleStep ELISA (ab227898), and leading colorimetric competitor. The Chemiluminescent SimpleStep ELISA kit shows increased sensitivity.

Sandwich ELISA - Human HIF-2-alpha ELISA Kit, Chemiluminescent (AB326404)
  • sELISA

Lab

Sandwich ELISA - Human HIF-2-alpha ELISA Kit, Chemiluminescent (AB326404)

Standard curve comparison between the Chemiluminescent Human HIF-2-alpha SimpleStep ELISA and colorimetric Human HIF-2-alpha SimpleStep ELISA (ab227898) in 1X Cell Extraction Buffer PTR. Background-subtracted data values (mean +/- SD) are graphed.

ELISA - Human HIF-2-alpha ELISA Kit, Chemiluminescent (AB326404)
  • ELISA

Supplier Data

ELISA - Human HIF-2-alpha ELISA Kit, Chemiluminescent (AB326404)

Interpolated concentrations of native HIF-2-alpha in DFO treated Human HeL1 cell extract sample based on a 250 µg/mL extract load.

HELA cells were untreated or treated with 250 µM DFO for 24 hrs then collected and extracted according to sample preparation protocol (see section 11.1). The concentrations of HIF-2-alpha were measured in duplicate and interpolated from the HIF-2-alpha standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD n=2). The mean HIF-2-alpha concentration was determined to be 520 pg/mL in DFO treated HELA cell extract and undetectable in untreated HELA cell extract.

ELISA - Human HIF-2-alpha ELISA Kit, Chemiluminescent (AB326404)
  • ELISA

Supplier Data

ELISA - Human HIF-2-alpha ELISA Kit, Chemiluminescent (AB326404)

Dose-dependent induction of HIF-2-alpha in HELA cells by deferoxamine (DFO).

HeLa cells were cultured in 96-well tissue culture plates and were either untreated (Vehicle Control) or exposed to varying doses of DFO for 24 hours (DFO treated). Cells were extracted directly in the culture plate by overlaying culture media with Cell Extraction Buffer PTR with Enhancer such that the final concentration was 1X Cell Extraction Buffer PTR. Extracts were applied to the HIF-2-alpha SimpleStep ELISA plate. Raw data and standard deviation is plotted from quadruplicate measurements.

Key facts

Detection method

Luminescent

Sample types

Cell culture extracts

Reacts with

Human

Assay type

Sandwich

Results type

Quantitative

Sensitivity

= 1.32 pg/mL

Range

1.83 - 4000 pg/mL

Assay time

55m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Human HIF-2-alpha ELISA Kit, Chemiluminescent (ab326404) is a rapid single-wash sandwich 55-min ELISA to measure Human HIF-2-alpha in Cell culture extracts with a sensitivity of 1.32 pg/mL.

How the assay works

Human HIF-2-alpha ELISA Kit, Chemiluminescent (ab326404) SimpleStep Ignite™ ELISA employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step during a 30 min only incubation, significantly reducing assay time. By utilizing a chemiluminscent detection method this improves the sensitivity and dynamic range when compared to traditional colorimetric ELISAs.

Assay Specificity

Our SimpleStep Ignite™ ELISA kits use recombinant monoclonal antibodies rigorously validated to ensure the highest level of consistency and reproducibility, improved sensitivity and specificity and ease of scalability and security of supply.
Please refer to our protocol booklet for more details.

Human HIF-2-alpha (ab326404) protocol summary

1. Mix : add samples / standards to the wells together with the capture and detector antibody cocktail. Incubate 30 min at room temperature
2. Wash
3. Add ChemiHRP development solution - incubate for 2 min
4. Read the results on a plate reader capable of luminescence detection

Design your own immunoassay

We offer the antibody pair used in this kit in a BSA and Azide-free format, ready for conjugation:

- Anti-HIF-2-alpha antibody [EPR19656-21] - BSA and Aside free (Capture) ab244690
- Anti-HIF-2-alpha antibody [EPR19656-33] - BSA and Aside free (Detector) ab244956
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Precision

[ { "reproducibilityType": "Intra", "sample": "HeLa Extract", "replicates": 5, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 2.6" }, { "reproducibilityType": "Inter", "sample": "HeLa Extract", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "= 4" } ]
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Recovery

[ { "sample": "Cell culture extracts", "range": "112 - 116 %", "average": "= 114" } ]
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What's included?

{ "values": { "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab326404-1x96Tests", "assetComponentDetails": [ { "size":"1 x 3 mL", "name":"ChemiHRP Reagent A", "number":"AB326404-CMP08", "productcode":"" }, { "size":"1 x 1 mL", "name":"50X Cell Extraction Enhancer Solution (ab193971)", "number":"AB326404-CMP06", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human HIF-2-alpha Capture Antibody", "number":"AB326404-CMP01", "productcode":"" }, { "size":"1 x 1 Unit", "name":"Plate Seal", "number":"AB326404-CMP11", "productcode":"" }, { "size":"1 x 1 Unit", "name":"SimpleStep Pre-Coated Black 96-Well Microplate", "number":"AB326404-CMP10", "productcode":"" }, { "size":"1 x 10 mL", "name":"5X Cell Extraction Buffer PTR (ab193970)", "number":"AB326404-CMP05", "productcode":"" }, { "size":"1 x 600 µL", "name":"10X Human HIF-2-alpha Detector Antibody", "number":"AB326404-CMP02", "productcode":"" }, { "size":"1 x 3 mL", "name":"ChemiHRP Reagent B", "number":"AB326404-CMP09", "productcode":"" }, { "size":"1 x 6 mL", "name":"Antibody Diluent CPR2", "number":"AB326404-CMP04", "productcode":"" }, { "size":"2 x 1 Vial", "name":"Human HIF-2-alpha Lyophilized Recombinant Protein", "number":"AB326404-CMP03", "productcode":"" }, { "size":"1 x 20 mL", "name":"10X Wash Buffer PT (ab206977)", "number":"AB326404-CMP07", "productcode":"" } ] } } }
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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C
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Product protocols

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Target data

Transcription factor involved in the induction of oxygen regulated genes. Heterodimerizes with ARNT; heterodimer binds to core DNA sequence 5'-TACGTG-3' within the hypoxia response element (HRE) of target gene promoters (By similarity). Regulates the vascular endothelial growth factor (VEGF) expression and seems to be implicated in the development of blood vessels and the tubular system of lung. May also play a role in the formation of the endothelium that gives rise to the blood brain barrier. Potent activator of the Tie-2 tyrosine kinase expression. Activation requires recruitment of transcriptional coactivators such as CREBBP and probably EP300. Interaction with redox regulatory protein APEX1 seems to activate CTAD (By similarity).
See full target information EPAS1
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websiteProtocolBooklet
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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