Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
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Species reactivity: | Human, Mouse, Rat |
Applications: | WB, IHC-P, IF-Cell, FC |
Clonality: | Monoclonal |
Clone number: | JE65-92 |
Form: | Liquid |
Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
Concentration: | 1ug/ul |
Purification: | Protein A affinity purified. |
Molecular weight: | Predicted band size: 41 kDa |
Isotype: | IgG |
Immunogen: | Recombinant protein within Human NFE2 aa 251-350 / 373. |
Positive control: | U-937 cell lysate, K-562 cell lysate, TF-1 cell lysate, HeLa cell lysate, HepG2 cell lysate, mouse testis tissue lysate, rat testis tissue lysate, human breast carcinoma tissue, SH-SY5Y, K-562. |
Subcellular location: | Cytoplasm, Nucleus. |
Recommended Dilutions:
WB IHC-P IF-Cell FC |
1:1000 1:200-1:1000 1:100 1:1000 |
Uniprot #: | SwissProt: Q16621 Human | Q07279 Mouse | Q6AYT2 Rat |
Alternative names: | Leucine zipper protein NF-E2 Nuclear factor, erythroid-derived 2 45 kDa subunit p45 NF-E2 |
Fig1:
Western blot analysis of NFE2 on different lysates with Rabbit anti-NFE2 antibody (HA721365) at 1/1,000 dilution. Lane 1: U-937 cell lysate (15 µg/Lane) Lane 2: K-562 cell lysate (15 µg/Lane) Lane 3: TF-1 cell lysate (15 µg/Lane) Lane 4: HeLa cell lysate (15 µg/Lane) Lane 5: HepG2 cell lysate (15 µg/Lane) Lane 6: Mouse testis tissue lysate (30µg/Lane) Lane 7: Rat testis tissue lysate (30µg/Lane) Predicted band size: 41 kDa Observed band size: 41 kDa Exposure time: 1 minute 20 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 (HA721365) 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. |
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Fig2:
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-NFE2 antibody (HA721365) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (HA721365) 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. |
Fig3:
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-NFE2 antibody (HA721365) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (HA721365) at 1/200 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. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-NFE2 antibody (HA721365) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (HA721365) at 1/200 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. |
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Fig5:
Immunocytochemistry analysis of K-562 cells labeling NFE2 with Rabbit anti-NFE2 antibody (HA721365) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-NFE2 antibody (HA721365) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
Fig6:
Flow cytometric analysis of K-562 cells labeling NFE2. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721365, 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). |