Keap1 ELISA Kits

NWLSS | Northwest Life Science Specialties, LLC.
NWK-KEAP1H1 $845.00 each

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The NWLSS™ Keap1 ELISA is intended for quantitative detection of Kelch-like ECH-associated protein 1 (KEAP1), also known as iNRF2, in either Human or Mouse species.
 
Variants of the NWLSS™ Keap1 ELISA:

  • Human [NWK-KEAP1H1]
  • Mouse [NWK-KEAP1M1]

Introduction

Nuclear factor, erythroid 2 like protein 2 (NFE2L2) commonly referred to as Nrf2 is an important protein in the regulation of oxidative and xenobiotic stress response. Kelch-like ECH-associated protein 1 (KEAP1), also known as inhibitor of Nrf2 (iNrf2), is equally important due to it being the primary regulator of Nrf2.

In the cytosol, KEAP1 normally binds to Nrf2 preventing it from translocating to the nucleus and targeting it for ubiquitination and subsequent degradation by proteasomes. However, KEAP1 is a cysteine rich protein that is sensitive to oxidative stress. When exposed to the appropriate stressor, oxidation of C151, C273 and C288 can produce a conformation change in Keap1 causing release of Nrf2 which then allows translocation of Nrf2 to the nucleus. Inside the nucleus, Nrf2 is free to participate in Nrf2/sMAF/ARE transcription pathways and is capable of activating more than three dozen xenobiotic and oxidative stress response genes. Many studies have shown a positive correlation between active Nrf2 and disease amelioration whereas others have shown heightened NRF2 and/or lower KEAP1 levels to increase cancer cell viability and resistance to chemotherapy.

In either role KEAP1 regulation of Nrf2 has emerged as a very important target for researchers trying to ascertain the full potential of the KEAP1/Nrf2/ARE pathway in terms of lessening the Deleterious effects of disease and aging.

Test Principle

The NWLSS™ Human KEAP1 ELISA is a sandwich format Enzyme-Linked Immunosorbent Assay (ELISA). The assay features a microtiter plate pre coated with a monoclonal antibody specific to human or mouse KEAP1. Briefly, Nrf2 in samples and standards is captured by the plate bound antibody and remaining liquid is removed. Next, a biotinylated antibody with specificity to KEAP1 is added and allowed to bind to KEAP1 captured in the previous step. After incubation, non-bound components are removed by washing. Avidin-HRP conjugate is then added and allowed to incubate.

After a subsequent thorough wash TMB-substrate solution is added to each well which produces blue coloration wherever KEAP1 is present. Finally, a sulfuric acid stop solution is added and the resulting yellow colored product is measured at 450nm. The amount of KEAP1 in the sample can be easily determined by direct comparison of unknown sample absorbance with known standard curve absorbance values generated in the assay.

For Research Use Only

Catalog Number: NWK-KEAP1H1
Format: 96 Well ELISA
Sample Requirements: Tissue Homogenates or Cell Lysates
Specificity: Human KEAP1
Sensitivity: 55 pg/mL
Standard Range: 0.156 ng/mL - 10 ng/mL
Storage and Stability: 6 months from the date of manufacture when individual components stored as specified.
Kit Contents: 1 12x8 Well Foil Pouch Microtiter plate precoated with anti-Human KEAP1
2 Vials KEAP1 Standard (Lyophilized)
1 Bottle Standard Diluent (20 mL)
1 Vial 100X Secondary Antibody (120 µL)
1 Bottle Secondary Antibody Diluent (12 mL)
1 Vial 100X Avidin-HRP Conjugate (120 µL)
1 Bottle Avidin-HRP Diluent (12 mL)
1 Bottle TMB Substrate (9 mL)
1 Bottle Stop Solution (6 mL)
1 Bottle 30X Wash Buffer (20 mL)
4 Adhesive Plate Covers
Catalog Number: NWK-KEAPM1
Format: 96 Well ELISA
Sample Requirements: Tissue Homogenates or Cell Lysates
Specificity: Mouse KEAP1
Sensitivity: 60 pg/mL
Standard Range: 0.156 ng/mL - 10 ng/mL
Storage and Stability: 6 months from the date of manufacture when individual components stored as specified.
Kit Contents: 1 12x8 Well Foil Pouch Microtiter plate precoated with anti-Mouse KEAP1
2 Vials KEAP1 Standard (Lyophilized)
1 Bottle Standard Diluent (20 mL)
1 Vial 100X Secondary Antibody (120 µL)
1 Bottle Secondary Antibody Diluent (12 mL)
1 Vial 100X Avidin-HRP Conjugate (120 µL)
1 Bottle Avidin-HRP Diluent (12 mL)
1 Bottle TMB Substrate (9 mL)
1 Bottle Stop Solution (6 mL)
1 Bottle 30X Wash Buffer (20 mL)
4 Adhesive Plate Covers

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