Keap1 Recombinant Rabbit Monoclonal Antibody [PSH0-90] - BSA and Azide free
Catalog# HA750670
Keap1 Recombinant Rabbit Monoclonal Antibody [PSH0-90] - BSA and Azide free
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WB
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IHC-P
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FC
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IP
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
Keap1 Recombinant Rabbit Monoclonal Antibody [PSH0-90] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human Keap1 aa 275-624 / 624.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, FC, IP
Target Molecular Weight
Predicted band size: 70 kDa
Positive Control
HepG2 cell lysate, HeLa cell lysate, MCF7 cell lysate, A431 cell lysate, Jurkat cell lysate, HEK-293 cell lysate, Daudi cell lysate, A549 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, mouse kidney tissue, rat kidney tissue, PC-12.
Conjugation
unconjugated
Clone Number
PSH0-90
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
Target
Function
Kelch-like ECH-associated protein 1 is a protein that in humans is encoded by the Keap1 gene. Keap1 has four discrete protein domains. The N-terminal Broad complex, Tramtrack and Bric-à-Brac (BTB) domain contains the Cys151 residue, which is one of the important cysteines in stress sensing. The intervening region (IVR) domain contains two critical cysteine residues, Cys273 and Cys288, which are a second group of cysteines important for stress sensing. A double glycine repeat (DGR) and C-terminal region (CTR) domains collaborate to form a β-propeller structure, which is where Keap1 interacts with Nrf2. Mutations in KEAP1 that result in loss-of-function are not linked to familial cancers, though they do predispose individuals to multinodular goiters. The proposed mechanism leading to goiter formation is that the redox stress experienced when the thyroid produces hormones selects for loss of heterozygosity of KEAP1, leading to the goiters.
Background References
1. Baird L et al. The Molecular Mechanisms Regulating the KEAP1-NRF2 Pathway. Mol Cell Biol. 2020 Jun
2. Koppula P et al. A targetable CoQ-FSP1 axis drives ferroptosis- and radiation-resistance in KEAP1 inactive lung cancers. Nat Commun. 2022 Apr
Subcellular Location
Cytoplasm, Nucleus.
Synonyms
Cytosolic inhibitor of Nrf2 antibody
INrf 2 antibody
INrf2 antibody
Keap 1 antibody
KEAP1 antibody
KEAP1_HUMAN antibody
Kelch like ECH associated protein 1 antibody
Kelch like family member 19 antibody
Kelch like protein 19 antibody
Kelch-like ECH-associated protein 1 antibody
ExpandImages
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This data was developed using HA721525, the same antibody clone in a different buffer formulation.
Western blot analysis of Keap1 on different lysates with Rabbit anti-Keap1 antibody (HA721525) at 1/1,000 dilution.
Lane 1: HepG2 cell lysate
Lane 2: HeLa cell lysate
Lane 3: MCF7 cell lysate
Lane 4: A431 cell lysate
Lane 5: Jurkat cell lysate
Lane 6: HEK-293 cell lysate
Lane 7: Daudi cell lysate
Lane 8: A549 cell lysate
Lane 9: NIH/3T3 cell lysate
Lane 10: PC-12 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 70 kDa
Observed band size: 55-70 kDa
Exposure time: 2 minutes;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721525) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature. -
This data was developed using HA721525, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Keap1 antibody (HA721525) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721525) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using HA721525, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Keap1 antibody (HA721525) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721525) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using HA721525, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of PC-12 cells labeling Keap1.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA721525, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
☑ Knockdown (KD)
This data was developed using HA721525, the same antibody clone in a different buffer formulation.
Western blot analysis of Keap1 on different lysates with Rabbit anti-Keap1 antibody (HA721525) at 1/1,000 dilution.
Lane 1: HepG2-si NT cell lysate
Lane 2: HepG2-si Keap1 cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 70 kDa
Observed band size: 55-70 kDa
Exposure time: 1 minute 55 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721525) at 1/1,000 dilution was used in 5% BSA at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
This data was developed using HA721525, the same antibody clone in a different buffer formulation.
Keap1 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA721525 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA721525 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input)
Lane 2: HA721525 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA721525 in HeLa cell lysate
Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 12 seconds; ECL: K1801
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"