N4BP1 Recombinant Rabbit Monoclonal Antibody [JE63-75]
cat.: HA722576
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Mouse, Rat |
| Applications: | WB, IF-Cell, IHC-P, FC |
| Clonality: | Monoclonal |
| Clone number: | JE63-75 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| 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: 99 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within mouse N4BP1 aa 251-500 / 893. |
| Positive control: | F9 cell lysate, C2C12 cell lysate, Neuro-2a cell lysate, PC-12 cell lysate, C2C12, mouse testis tissue, rat testis tissue. |
| Subcellular location: | Cytoplasm, cytosol, Nucleus, nucleolus, PML body. |
| Recommended Dilutions:
WB IF-Cell IHC-P FC |
1:1,000 1:100 1:1,000 1:1,000 |
| Uniprot #: | SwissProt: Q6A037 Mouse Entrez Gene: 291921 Rat |
| Alternative names: | FLJ31821 KIAA0615 MGC176730 N4BP1 N4BP1_HUMAN NEDD4 binding protein 1 NEDD4-binding protein 1 |
Images
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Fig1:
Western blot analysis of N4BP1 on different lysates with Rabbit anti-N4BP1 antibody (HA722576) at 1/1,000 dilution. Lane 1: F9 cell lysate Lane 2: C2C12 cell lysate Lane 3: Neuro-2a cell lysate Lane 4: PC-12 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 99 kDa Observed band size: 110 kDa Exposure time: 59 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 (HA722576) 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. |
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Fig2:
Immunocytochemistry analysis of C2C12 cells labeling N4BP1 with Rabbit anti-N4BP1 antibody (HA722576) 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-N4BP1 antibody (HA722576) 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 (M1305-2, 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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Fig3:
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-N4BP1 antibody (HA722576) 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 (HA722576) 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. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-N4BP1 antibody (HA722576) 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 (HA722576) 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. |
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Fig5:
Flow cytometric analysis of C2C12 cells labeling N4BP1. Cells were fixed and permeabilized. Then stained with the primary antibody (HA722576, 1/1,000) (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). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.