Nrf2 Recombinant Rabbit Monoclonal Antibody [PSH11-20]
cat.: HA723302
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, ChIP |
| Clonality: | Monoclonal |
| Clone number: | PSH11-20 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 68 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human Nrf2 aa 51-350. |
| Positive control: | MEF treated with 10μM MG-132 for 8 hours cell lysate, HeLa treated with 2μM MG-132 for 18 hours cell lysate, C6 treated with 25μM MG-132 for 4 hours cell lysate, MEF cells treated with 2μM MG-132 for 18 hours, HCT 116 cells treated with 25μM MG-132 for 4 hours. |
| Subcellular location: | Cytoplasm, cytosol, Nucleus. |
| Recommended Dilutions:
WB IF-Cell ChIP |
1:500-1:2,000 1:100 Use 0.5~2 μg for 25 μg of chromatin. |
| Uniprot #: | SwissProt: Q16236 Human | Q60795 Mouse | O54968 Rat |
| Alternative names: | erythroid derived 2 HEBP1 like 2 NF E2 related factor 2 NF-E2-related factor 2 NF2L2_HUMAN NFE2 related factor 2 NFE2-related factor 2 Nfe2l2 Nrf 2 NRF2 Nuclear factor (erythroid derived 2) like 2 Nuclear factor nuclear factor erythroid 2 like 2 Nuclear factor erythroid 2 related factor 2 Nuclear factor erythroid 2-related factor 2 Nuclear factor erythroid derived 2 like 2 |
Images
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Fig1:
Western blot analysis of Nrf2 on different lysates with Rabbit anti-Nrf2 antibody (HA723302) at 1/2,000 dilution. Lane 1: MEF cell lysate Lane 2: MEF treated with 10μM MG-132 for 8 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 68 kDa Observed band size: 100 kDa Exposure time: 14 seconds; ECL: K1801; 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 (HA723302) at 1/2,000 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. |
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Fig2:
Western blot analysis of Nrf2 on different lysates with Rabbit anti-Nrf2 antibody (HA723302) at 1/2,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 2μM MG-132 for 18 hours cell lysate Lane 3: C6 cell lysate Lane 4: C6 treated with 25μM MG-132 for 4 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 68 kDa Observed band size: 100 kDa Exposure time: 2 minutes; ECL: K1801; 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 (HA723302) at 1/2,000 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. |
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Fig3:
Immunocytochemistry analysis of MEF cells untreated / treated with 2μM MG-132 for 18 hours labeling Nrf2 with Rabbit anti-Nrf2 antibody (HA723302) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 (HA723302) 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. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Immunocytochemistry analysis of HCT 116 cells untreated / treated with 25μM MG-132 for 4 hours labeling Nrf2 with Rabbit anti-Nrf2 antibody (HA723302) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 (HA723302) 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. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig5: Chromatin immunoprecipitations were performed with cross-linked chromatin from MEF cells treated with 10μM MG-132 for 8 hours with Nrf2 (HA723302) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. |
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