Anti-KEAP1 Antibody Picoband™

Product Name

Anti-KEAP1 Antibody Picoband™

View all KEAP1 Antibodies

SKU/Catalog Number

A00514-3

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-KEAP1 Antibody Picoband™ catalog # A00514-3. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-KEAP1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00514-3)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human KEAP1 recombinant protein (Position: K84-K312).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A00514-3 is reactive to KEAP1 in Human, Mouse, Rat

Applications

A00514-3 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

66-72 kDa

Calculated molecular weight

69.666kDa

Background of KEAP1

KEAP1 (KELCH-LIKE ECH-ASSOCIATED PROTEIN 1), is a protein that in humans is encoded by the Keap1 gene. The KIAA0132 gene is mapped on 19p13.2. Keap1 contains a central BTB/POZ domain and a C-terminal double glycine repeat (DGR), or Kelch, module. Keap1 has been shown to interact with Nrf2, a master regulator of the antioxidant response, which is important for the amelioration of oxidative stress. In the presence of the electrophilic agent diethylmalate, Nrf2 activity is released from Keap1 and Nrf2 translocate to the nucleus. Under quiescent conditions, Nrf2 is anchored in the cytoplasm through binding to Keap1, which, in turn, facilitates the ubiquitination and subsequent proteolysis of Nrf2. Because Nrf2 activation leads to a coordinated antioxidant and anti-inflammatory response, and Keap1 represses Nrf2 activation, Keap1 has become a very attractive drug target.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 μg/ml, Human, Mouse, Rat
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human, Rat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of KEAP1 using anti-KEAP1 antibody (A00514-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human 293T whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: rat C6 whole cell lysates,
Lane 6: mouse Neuro-2a whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-KEAP1 antigen affinity purified polyclonal antibody (Catalog # A00514-3) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for KEAP1 at approximately 66-72 kDa. The expected band size for KEAP1 is at 70 kDa.

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Figure 2. IF analysis of KEAP1 using anti-KEAP1 antibody (A00514-3).
KEAP1 was detected in an immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-KEAP1 Antibody (A00514-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of U251 cells using anti-KEAP1 antibody (A00514-3).
Overlay histogram showing U251 cells stained with A00514-3 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-KEAP1 Antibody (A00514-3, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For KEAP1 (Source: Uniprot.org, NCBI)

Alternative Names

INrf2; INrf2MGC9454; Keap1; kelch-like ECH-associated protein 1; Kelch-like protein 19; KIAA0132MGC4407; KLHL19Cytosolic inhibitor of Nrf2; MGC10630; MGC1114; MGC20887 KEAP1 INrf2, KLHL19 kelch like ECH associated protein 1 kelch-like ECH-associated protein 1|KEAP1 delta C|cytosolic inhibitor of Nrf2|kelch-like family member 19|kelch-like protein 19

For more info on KEAP1, check out the KEAP1 Infographic

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In this infographic, you will see the following information for KEAP1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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