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Nrf2 Recombinant Rabbit Monoclonal Antibody [JE30-14]

Usd: 385

Specification

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Catalog# HA721432

Nrf2 Recombinant Rabbit Monoclonal Antibody [JE30-14]

Application

  • WB

  • IF-Cell

Reactivity

  • Human

  • Mouse

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Nrf2 Recombinant Rabbit Monoclonal Antibody [JE30-14]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Nrf2 aa 550-605.

Species Reactivity

Human, Mouse

Validated Applications

WB, IF-Cell

Molecular Weight

Predicted band size: 68 kDa

Positive Control

HeLa whole cell lysate, HeLa treated with 2μM MG-132 for 18 hours whole cell lysate, HCT 116 whole cell lysate, HCT 116 treated with 25μM MG-132 for 4 hours whole cell lysate, NIH/3T3 treated with 10μM MG-132 for 8 hours cell lysate, HCT 116.

Conjugation

unconjugated

Clone Number

JE30-14

RRID

AB_3073494

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

Target

Function

Nuclear factor erythroid 2-related factor 2 (NRF2), also known as nuclear factor erythroid-derived 2-like 2, is a transcription factor that in humans is encoded by the NFE2L2 gene. NRF2 is a basic leucine zipper (bZIP) protein that may regulate the expression of antioxidant proteins that protect against oxidative damage triggered by injury and inflammation, according to preliminary research.[6] In vitro, NRF2 binds to antioxidant response elements (AREs) in the promoter regions of genes encoding cytoprotective proteins. NRF2 induces the expression of heme oxygenase 1 in vitro leading to an increase in phase II enzymes. NRF2 also inhibits the NLRP3 inflammasome. NRF2 appears to participate in a complex regulatory network and performs a pleiotropic role in the regulation of metabolism, inflammation, autophagy, proteostasis, mitochondrial physiology, and immune responses.

Background References

1. Masuda Y et al. The effect of Nrf2 pathway activation on human pancreatic islet cells. PLoS One 10:e0131012 (2015).

2. Stachel I et al. Modulation of nuclear factor E2-related factor-2 (Nrf2) activation by the stress response gene immediate early response-3 (IER3) in colonic epithelial cells: a novel mechanism of cellular adaption to inflammatory stress. J Biol Chem 289:1917-29 (2014).

Sequence Similarity

Belongs to the bZIP family. CNC subfamily.

Tissue Specificity

Widely expressed. Highest expression in adult muscle, kidney, lung, liver and in fetal muscle.

Post-translational Modification

Ubiquitinated in the cytoplasm by the BCR(KEAP1) E3 ubiquitin ligase complex leading to its degradation. In response to oxidative stress, electrophile metabolites, such as sulforaphane, modify KEAP1, leading to inhibit activity of the BCR(KEAP1) complex, promoting NFE2L2/NRF2 nuclear accumulation and activity. In response to autophagy, the BCR(KEAP1) complex is inactivated (By similarity).; Phosphorylation of Ser-40 by PKC in response to oxidative stress dissociates NFE2L2 from its cytoplasmic inhibitor KEAP1, promoting its translocation into the nucleus.; Acetylation at Lys-596 and Lys-599 increases nuclear localization whereas deacetylation by SIRT1 enhances cytoplasmic presence.; Glycation impairs transcription factor activity by preventing heterodimerization with small Maf proteins. Deglycation by FN3K restores activity.

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: Q16236 Human

SwissProt: Q60795 Mouse

Synonyms

erythroid derived 2 antibody

HEBP1 antibody

like 2 antibody

NF E2 related factor 2 antibody

NF-E2-related factor 2 antibody

NF2L2_HUMAN antibody

NFE2 related factor 2 antibody

NFE2-related factor 2 antibody

Nfe2l2 antibody

Nrf 2 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Nrf2 on different lysates with Rabbit anti-Nrf2 antibody (<a href="/products/HA721432" style="font-weight: bold;text-decoration: underline;">HA721432</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa whole cell lysate<br />Lane 2: HeLa treated with 2μM MG-132 for 18 hours whole cell lysate<br />Lane 3: HCT 116 whole cell lysate<br />Lane 4: HCT 116 treated with 25μM MG-132 for 4 hours whole cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 68 kDa<br />Observed band size: 110 kDa<br /><br />Exposure time: 3 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721432" style="font-weight: bold;text-decoration: underline;">HA721432</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Nrf2 on different lysates with Rabbit anti-Nrf2 antibody (HA721432) at 1/1,000 dilution.

    Lane 1: HeLa whole cell lysate
    Lane 2: HeLa treated with 2μM MG-132 for 18 hours whole cell lysate
    Lane 3: HCT 116 whole cell lysate
    Lane 4: HCT 116 treated with 25μM MG-132 for 4 hours whole cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 68 kDa
    Observed band size: 110 kDa

    Exposure time: 3 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 (HA721432) 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Nrf2 on different lysates with Rabbit anti-Nrf2 antibody (<a href="/products/HA721432" style="font-weight: bold;text-decoration: underline;">HA721432</a>) at 1/1,000 dilution.<br /><br />Lane 1: NIH/3T3 cell lysate<br />Lane 2: NIH/3T3 treated with 10μM MG-132 for 8 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 68 kDa<br />Observed band size: 110 kDa<br /><br />Exposure time: 3 minutes;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721432" style="font-weight: bold;text-decoration: underline;">HA721432</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Nrf2 on different lysates with Rabbit anti-Nrf2 antibody (HA721432) at 1/1,000 dilution.

    Lane 1: NIH/3T3 cell lysate
    Lane 2: NIH/3T3 treated with 10μM MG-132 for 8 hours cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 68 kDa
    Observed band size: 110 kDa

    Exposure time: 3 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 (HA721432) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HCT 116 cells labeling Nrf2 with Rabbit anti-Nrf2 antibody (<a href="/products/HA721432" style="font-weight: bold;text-decoration: underline;">HA721432</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Nrf2 antibody (<a href="/products/HA721432" style="font-weight: bold;text-decoration: underline;">HA721432</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of HCT 116 cells labeling Nrf2 with Rabbit anti-Nrf2 antibody (HA721432) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Nrf2 antibody (HA721432) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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