Keap1 Antibody (333116)

R&D Systems | Catalog # MAB3024

R&D Systems

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Mouse, Rat

Applications

Validated:

Knockout Validated, Western Blot, Simple Western

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 333116
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Product Specifications

Immunogen

E. coli-derived recombinant human Keap1
Ala90-Ile250
Accession # Q14145

Specificity

Detects human, mouse and rat Keap1 in Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Keap1 Antibody (333116)

Detection of Human/Mouse/Rat Keap1 antibody by Western Blot.

Detection of Human/Mouse/Rat Keap1 by Western Blot.

Western blot shows lysates of Jurkat human acute T cell leukemia cell line, Raji human Burkitt's lymphoma cell line, Neuro-2A mouse neuroblastoma cell line, and L6 rat myoblast cell line. PVDF membrane was probed with 1 µg/mL of Human/Mouse/Rat Keap1 Monoclonal Antibody (Catalog # MAB3024) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Keap1 at approximately 66 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 4.
Detection of Human Keap1 antibody by Simple WesternTM.

Detection of Human Keap1 by Simple WesternTM.

Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for Keap1 at approximately 75 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human/Mouse/Rat Keap1 Monoclonal Antibody (Catalog # MAB3024). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.
Western Blot Shows Human Keap1 Antibody Specificity by Using Knockout Cell Line.

Western Blot Shows Human Keap1 Specificity by Using Knockout Cell Line.

Western blot shows lysates of Jurkat human acute T cell leukemia parental cell line and Keap1 Jurkat cell line (KO). PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human/Mouse/Rat Keap1 Monoclonal Antibody (Catalog # MAB3024) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Keap1 at approximately 66 kDa (as indicated) in the parental Jurkat cell line, but is not detectable in knockout Jurkat cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Keap1 by Western Blot

Detection of Keap1 by Western Blot

Effect of RSO on Nrf2-Keap1 pathway of LPS-induced RAW 264.7 cells. Relative mRNA expression of Nrf2 (A), HO-1 (B), NQO1 (C), and Keap1 (D) genes normalized to beta -actin and expressed as fold change of the control group, n = 3. (E) The protein levels of Nrf2 and Keap1 were analyzed by immunoblotting with beta -actin as the internal reference protein for normalization, n = 3. Relative protein expression of Nrf2 (F) and Keap1 (G) were expressed as fold change of the LPS-induced group. ** p < 0.01 vs control group; # p < 0.05, ## p < 0.01 vs LPS-induced group. Image collected and cropped by CiteAb from the following open publication (https://www.mdpi.com/2072-6643/14/7/1349), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Keap1 by Western Blot

Detection of Keap1 by Western Blot

MeHg induced the expression of antioxidant enzymes in C8-D1A astrocytic cells. Astrocytic C8-D1A cells were treated with MeHg (0, 0.1, 0.5, 1, 5, 10, or 20 µM). Western blot analysis was used to measure changes in the antioxidant proteins Nrf2 (A), KEAP1 (B), HO-1 (C), and SOD2 (D) in C8-D1A cells exposed to 10 µM MeHg for 30 min, 1, 3, 6, or 24 h. (E) shows representative densitometry images. Hmox1 (F), Nqo1 (G), Sod2 (H), and Slc7a11 (I) gene expression were measured using qPCR in cells exposed to 10 µM MeHg for 1, 3, or 6 h. Data are presented as mean ± SD. Statistical significance was determined using one-way ANOVA followed by Bonferroni’s post-hoc analysis, or with the Kruskal-Wallis test followed by Dunn’s post hoc test, adjusted by Bonferroni correction when normality was not achieved. p < 0.05 was considered statistically significant, * denotes a significant difference Image collected and cropped by CiteAb from the following open publication (https://link.springer.com/10.1007/s11064-025-04507-7), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Keap1 by Western Blot

Detection of Keap1 by Western Blot

NRF2 expression is positively associated with malignancy in HNSCC. (A, C, D) Representative IHC staining of NRF2 in the specimens obtained from HNSCC patients. Brown staining indicates NRF2 positivity. The cell nuclei and background were counterstained with hematoxylin (blue coloration). (A) NRF2 protein expression in HNSCC specimens with different clinicopathological stages. Magnification, 100× (upper panel) and 400× (lower panel). (B) The correlation between the level of nuclear and cytosolic expression of NRF2 and tumor stage was estimated using the immunoreactivity scoring system (IRS). The IRS was calculated by multiplying the intensity of NRF2 staining (scale, 0-3) by the percentage of positive cells (4, > 80%; 3, 51-80%; 2, 10-50%; 1, < 10%; 0, 0%), which resulted in values ranging from 0 to 12. Groups with different letters are significant different (p < 0.05) from each other. (C) NRF2 protein expression was positively correlated with the degree of tumor cell differentiation in human head and neck squamous cell carcinoma. Magnification, 50× (upper panel) and 400× (lower panel). (D) NRF2 expression was highest in the invasive carcinoma nests. Magnification 50×. (E) Levels of NRF2 and NRF2 downstream targets in dysplasia oral keratinocyte (DOK) and HNSCC cell lines. (F) Motility of DOK and various HNSCC cell lines via trans-well migration assay. * p < 0.05; ** p < 0.01; *** p < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33859744), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Keap1 Antibody (333116)

Application
Recommended Usage

Knockout Validated

Keap1 is specifically detected in Jurkat human acute T cell leukemia parental cell line but is not detectable in Keap1 knockout Jurkat cell line.

Simple Western

20 µg/mL
Sample: Jurkat human acute T cell leukemia cell line

Western Blot

1 µg/mL
Sample: Jurkat human acute T cell leukemia cell line, Raji human Burkitt's lymphoma cell line, Neuro-2A mouse neuroblastoma cell line, and L6 rat myoblast cell line

Reviewed Applications

Read 1 review rated 4 using MAB3024 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.


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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Keap1

Kelch-like ECH-associated protein 1 (Keap1) is a 624 amino acid, 69 kDa protein which interacts with the transcription factor NF-E2-related factor 2 (Nrf2). Keap1 represses Nrf2 function by sequestering Nrf2 in the cytoplasm. Keap1 contains an N-terminal BTB domain and six C-terminal KELCH domains (aa 327‑611) that interact with Nrf2. Dissociation of the two proteins in response to redox-sensitive cell stress is followed by the translocation of Nrf2 to the nucleus and transcription of detoxifying/oxidative stress enzyme genes.

Long Name

Kelch-like ECH-associated Protein 1

Alternate Names

INrf2

Entrez Gene IDs

9817 (Human)

Gene Symbol

KEAP1

UniProt

Additional Keap1 Products

Product Documents for Keap1 Antibody (333116)

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Keap1 Antibody (333116)

For research use only

Citations for Keap1 Antibody (333116)

Customer Reviews for Keap1 Antibody (333116) (1)

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  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 22836768
    Species: Human
    Verified Customer | Posted 02/19/2015

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