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Western Blot (WB) (Figure 1. Western blot analysis of NFE2L2 using anti-NFE2L2 antibody (MBS1753271).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 30ug of sample under reducing conditions.Lane 1: human HepG2 whole cell lysatesLane 2: human SW620 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-NFE2L2 antigen affinity purified polyclonal antibody (Catalog # MBS1753271) at 0. 5 μg/mL overnight at 4 degree 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 # MBS176460) with Tanon 5200 system. A specific band was detected for NFE2L2 at approximately 100KD. The expected band size for NFE2L2 is at 100KD. )

Rabbit anti-Human NFE2L2 Polyclonal Antibody | anti-NFE2L2 antibody

Anti-NFE2L2 Antibody

Gene Names
NFE2L2; NRF2
Reactivity
Human
Applications
Western Blot, Immunohistochemistry, Immunocytochemistry, Immunofluorescence, Flow Cytometry, Functional Assay, ELISA
Purity
Immunogen affinity purified.
Synonyms
NFE2L2; Polyclonal Antibody; Anti-NFE2L2 Antibody; NRF2; Nuclear factor erythroid 2-related factor 2; NF-E2-related factor 2; NFE2-related factor 2; Nrf-2; HEBP1; Nuclear factor; erythroid derived 2; like 2; nuclear factor; erythroid 2 like 2; anti-NFE2L2 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Rabbit IgG polyclonal antibody for NFE2L2 detection.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.
Applicable Applications for anti-NFE2L2 antibody
Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA)
Application Notes
WB: 0.25-0.5ug/ml|Human|
IHC-P: 2-5ug/ml|Human|
ICC/IF: 5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Direct ELISA: 0.1-0.5ug/ml|Human|
Immunogen
E Coli-derived human NFE2L2 recombinant protein (Position: R34-N605).
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Recommended Detection Systems
Recommended Detection Systems
Preparation and Storage
Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

Western Blot (WB)

(Figure 1. Western blot analysis of NFE2L2 using anti-NFE2L2 antibody (MBS1753271).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 30ug of sample under reducing conditions.Lane 1: human HepG2 whole cell lysatesLane 2: human SW620 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-NFE2L2 antigen affinity purified polyclonal antibody (Catalog # MBS1753271) at 0. 5 μg/mL overnight at 4 degree 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 # MBS176460) with Tanon 5200 system. A specific band was detected for NFE2L2 at approximately 100KD. The expected band size for NFE2L2 is at 100KD. )

Western Blot (WB) (Figure 1. Western blot analysis of NFE2L2 using anti-NFE2L2 antibody (MBS1753271).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 30ug of sample under reducing conditions.Lane 1: human HepG2 whole cell lysatesLane 2: human SW620 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-NFE2L2 antigen affinity purified polyclonal antibody (Catalog # MBS1753271) at 0. 5 μg/mL overnight at 4 degree 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 # MBS176460) with Tanon 5200 system. A specific band was detected for NFE2L2 at approximately 100KD. The expected band size for NFE2L2 is at 100KD. )

Immunohistochemistry (IHC)

(Figure 2. IHC analysis of NFE2L2 using anti-NFE2L2 antibody (MBS1753271).NFE2L2 was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NFE2L2 Antibody (MBS1753271) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )

Immunohistochemistry (IHC) (Figure 2. IHC analysis of NFE2L2 using anti-NFE2L2 antibody (MBS1753271).NFE2L2 was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-NFE2L2 Antibody (MBS1753271) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )

Immunofluorescence (IF)

(Figure 3. IF analysis of NFE2L2 using anti- NFE2L2 antibody (MBS1753271).NFE2L2 was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti- NFE2L2 Antibody (MBS1753271) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )

Immunofluorescence (IF) (Figure 3. IF analysis of NFE2L2 using anti- NFE2L2 antibody (MBS1753271).NFE2L2 was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti- NFE2L2 Antibody (MBS1753271) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )

Flow Cytometry (FC/FACS)

(Figure 4. Flow Cytometry analysis of 293T cells using anti-NFE2L2 antibody (MBS1753271).Overlay histogram showing 293T cells stained with MBS1753271 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NFE2L2 Antibody (MBS1753271, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Flow Cytometry (FC/FACS) (Figure 4. Flow Cytometry analysis of 293T cells using anti-NFE2L2 antibody (MBS1753271).Overlay histogram showing 293T cells stained with MBS1753271 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NFE2L2 Antibody (MBS1753271, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Related Product Information for anti-NFE2L2 antibody
NFE2L2(nuclear factor (erythroid-derived 2)-like 2) also known as NRF2 or NFE2-RELATED TRANSCRIPTION FACTOR 2, is a transcription factor that in humans is encoded by the NFE2L2 gene. NFE2, NFE2L1, and NFE2L2 comprise a family of human genes encoding basic leucine zipper (bZIP) transcription factors. NFE2L2 induces the expression of various genes including those that encode for several antioxidant enzymes, and it may play a physiological role in the regulation of oxidative stress. The NFE2L2 gene is located on 2q31. 2. The identification of somatic mutations that disrupt the NRF2-KEAP1 interaction to stabilize NRF2 and increase the constitutive transcription of NRF2 target genes indicated that enhanced reactive oxygen species (ROS) detoxification and additional NRF2 functions may in fact be tumorigenic. Oncogene-directed increased expression of Nrf2 is a mechanism for the activation of the Nrf2 antioxidant program evident in primary cells and tissues of mice expressing KRas(G12D) and BRaf(V619E), and in human pancreatic cancer.
References
1. Chan, J. Y., Cheung, M. -C., Moi, P., Chan, K., Kan, Y. W. Chromosomal localization of the human NF-E2 family of bZIP transcription factors by fluorescence in situ hybridization. Hum. Genet. 95: 265-269, 1995.
2. He, C. H., Gong, P., Hu, B., Stewart, D., Choi, M. E., Choi, A. M. K., Alam, J. Identification of activating transcription factor 4 (ATF4) as an Nrf2-interacting protein: implication for heme oxygenase-1 gene regulation. J. Biol. Chem. 276: 20858-20865, 2001.
3. Lee, J. -M., Chan, K., Kan, Y. W., Johnson, J. A. Targeted disruption of Nrf2 causes regenerative immune-mediated hemolytic anemia. Proc. Nat. Acad. Sci. 101: 9751-9756, 2004.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
67,827 Da
NCBI Official Full Name
nuclear factor erythroid 2-related factor 2 isoform 2
NCBI Official Synonym Full Names
nuclear factor (erythroid-derived 2)-like 2
NCBI Official Symbol
NFE2L2
NCBI Official Synonym Symbols
NRF2
NCBI Protein Information
nuclear factor erythroid 2-related factor 2; HEBP1; NFE2-related factor 2; NF-E2-related factor 2; nuclear factor erythroid-derived 2-like 2; nuclear factor, erythroid derived 2, like 2
UniProt Protein Name
Nuclear factor erythroid 2-related factor 2
UniProt Gene Name
NFE2L2
UniProt Synonym Gene Names
NRF2; NF-E2-related factor 2
UniProt Entry Name
NF2L2_HUMAN

NCBI Description

This gene encodes a transcription factor which is a member of a small family of basic leucine zipper (bZIP) proteins. The encoded transcription factor regulates genes which contain antioxidant response elements (ARE) in their promoters; many of these genes encode proteins involved in response to injury and inflammation which includes the production of free radicals. Multiple transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jan 2012]

Uniprot Description

NRF2: a transcription factor that regulates basal expression and antioxidant induction of a transcription factor for NAD(P)H:quinone oxidoreductase-1 (NQO1) and other detoxifying genes. Targeted for proteasomal degradation by INrf2. Oxidative stress causes its phosphorylation and release from INrf2.

Protein type: DNA-binding; Transcription factor

Chromosomal Location of Human Ortholog: 2q31

Cellular Component: nucleoplasm; centrosome; cytoplasm; plasma membrane; nucleus; chromatin; cytosol

Molecular Function: protein domain specific binding; protein binding; DNA binding; transcription factor activity

Biological Process: transcription from RNA polymerase II promoter; proteasomal ubiquitin-dependent protein catabolic process; positive regulation of blood coagulation; unfolded protein response; regulation of embryonic development; proteasomal ubiquitin-independent protein catabolic process; protein ubiquitination; positive regulation of transcription from RNA polymerase II promoter

Research Articles on NFE2L2

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Product Notes

The NFE2L2 nfe2l2 (Catalog #AAA1753271) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-NFE2L2 Antibody reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's NFE2L2 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Human| IHC-P: 2-5ug/ml|Human| ICC/IF: 5ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human| Direct ELISA: 0.1-0.5ug/ml|Human|. Researchers should empirically determine the suitability of the NFE2L2 nfe2l2 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "NFE2L2, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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