Anti-Nrf2 antibody (ab137550)
Key features and details
- Rabbit polyclonal to Nrf2
- Suitable for: IP, ChIP, WB, ICC/IF, IHC-P
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Nrf2 antibody
See all Nrf2 primary antibodies -
Description
Rabbit polyclonal to Nrf2 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ChIP HumanICC/IF HumanIHC-P HumanIP HumanWB Human
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Immunogen
Recombinant fragment corresponding to Human Nrf2 aa 108-413 (internal sequence).
Database link: Q16236 -
Positive control
- WB: MDA-MB-231 nuclear extract treated with 30 µM tBHQ for 4 hours (+). NRF2-transfected (+, including 3xFlag-tag) HEK-293T whole cell extracts. ICC/IF: HeLa cells. IHC: Human breast carcinoma tissue. ChIP: HepG2 chromatin extract. IP: HepG2 whole cell extracts.
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General notes
NRF2 is bound to ubiquitin when it is the nucleus so MW would be higher than cytoplasm.
Please note:
Nrf2 antibody (ab137550) detects no signal in most untreated samples for WB. Stimuli treated samples are recommended. Nrf2 expression is stimulated by oxidative stress, electrophiles and chemical activators (PMID: 25761198, PMID: 27638861 and PMID: 28587109).
WB protocol info included in legend for MDA-MB-231 nuclear extract treated with 30 µM tBHQ for 4 hours (+).
Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).
See other anti-rabbit secondary antibodies that can be used with this antibody.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.00
Preservative: 0.025% Proclin 300
Constituents: 78% PBS, 1% BSA, 20% Glycerol (glycerin, glycerine) -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nrf2 antibody (ab137550)
Paraffin embedded human breast carcinoma tissue stained for NRF2 using ab137550 at 1/500 dilution in immunohistochemical analysis.
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Western blot - Anti-Nrf2 antibody (ab137550)All lanes : Anti-Nrf2 antibody (ab137550) at 1/1000 dilution
Lane 1 : Untreated (-) MDA-MB-231 nuclear extract
Lane 2 : MDA-MB-231 nuclear extract treated with 30 µM tBHQ for 4 hours (+)
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Rabbit IgG antibody (HRP) at 1/10000 dilution
Developed using the ECL technique.
Observed band size: 110 kDa why is the actual band size different from the predicted?7.5% gel.
Running condition: 80V, 15min; 140V, 40min.
Transfer condition:Semi-dry, 18 V, 60min (Nitrocellulose membrane)
Blocking condition: 5% non-fat milk in TBST, RT, 60min.
Primary antibody incubation: 4°C overnight.
Secondary antibody incubation: Room temperature for 1 hour.
Washing condition: 5 ml TBST, 4 x 5min.
ECL detection.
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Immunocytochemistry/ Immunofluorescence - Anti-Nrf2 antibody (ab137550)ab137550 staining Nrf2 in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% paraformaldehyde. Samples were incubated with primary antibody diluted at 1/500 (green). Phalloidin, a cytoskeleton marker, diluted at 1/100 (red) was used.
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ChIP - Anti-Nrf2 antibody (ab137550)ChIP was performed with HepG2 chromatin extract and 5 μg of either normal rabbit IgG or Anti-Nrf2 antibody (ab137550). The precipitated DNA was detected by PCR with primer set targeting to GCLC gene locus.
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Immunoprecipitation - Anti-Nrf2 antibody (ab137550)ab137550 at 5 μg immunoprecipitating Nrf2 in HepG2 whole cell extracts.
Lane 1 (-): Ctrl IgG instead of ab137550 in HepG2 whole cell extracts.
Lane 2 (+): ab137550 + HepG2 whole cell extracts.For western blotting an anti-Rabbit IgG was used as a secondary reagent.
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Western blot - Anti-Nrf2 antibody (ab137550)All lanes : Anti-Nrf2 antibody (ab137550) at 1/1000 dilution
Lane 1 : Non-transfected (-) HEK-293T whole cell extracts
Lane 2 : NRF2-transfected (+, including 3xFlag-tag) HEK-293T whole cell extracts
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : HRP-conjugated anti-rabbit IgG antibody
Observed band size: 110 kDa why is the actual band size different from the predicted?5% SDS-PAGE
