SP1 Recombinant Rabbit Monoclonal Antibody [JF0950]
cat.: ET1702-02
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IF-Tissue, IHC-P, FC, ChIP, IP |
| Clonality: | Monoclonal |
| Clone number: | JF0950 |
| 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: 81 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within Human SP1 aa 544-588 / 785. |
| Positive control: | HeLa cell lysate, 293T cell lysate, Jurkat cell lysate, U-2 OS cell lysate, COS-1 cell lysate, HeLa, human breast cancer tissue, human colon cancer tissue, human lung tissue, mouse lung tissue, rat lung tissue, MCF-7. |
| Subcellular location: | Nucleus, Cytoplasm. |
| Recommended Dilutions:
WB IF-Cell IF-Tissue IHC-P FC ChIP IP |
1:1,000-1:2,000 1:100-1:500 1:100-1:500 1:200-1:1,000 1:500-1:1,000 Use 0.5~2 μg for 25 μg of chromatin. 1-2μg/sample |
| Uniprot #: | SwissProt: P08047 Human | O89090 Mouse | Q01714 Rat |
| Alternative names: | SP 1 SP1 Sp1 transcription factor SP1_HUMAN Specificity protein 1 Transcription factor Sp1 TSFP 1 TSFP1 |
Images
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Fig1:
Western blot analysis of SP1 on different lysates with Rabbit anti-SP1 antibody (ET1702-02) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: 293T cell lysate Lane 3: Jurkat cell lysate Lane 4: U-2 OS cell lysate Lane 5: COS-1 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 81 kDa Observed band size: 81 kDa Exposure time: 4 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 (ET1702-02) 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 HeLa cells labeling SP1 with Rabbit anti-SP1 antibody (ET1702-02) at 1/200 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-SP1 antibody (ET1702-02) at 1/200 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. |
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Fig3:
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-SP1 antibody (ET1702-02) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (ET1702-02) 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 human colon cancer tissue with Rabbit anti-SP1 antibody (ET1702-02) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (ET1702-02) 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:
Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-SP1 antibody (ET1702-02) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (ET1702-02) 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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Fig6:
Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Rabbit anti-SP1 antibody (ET1702-02) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (ET1702-02) 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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Fig7:
Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-SP1 antibody (ET1702-02) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (ET1702-02) 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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Fig8:
Flow cytometric analysis of MCF-7 cells labeling SP1. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-02, 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). |
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Fig9: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with SP1 (ET1702-02) 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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Fig10:
SP1 was immunoprecipitated from 0.2 mg HeLa cell lysate with ET1702-02 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1702-02 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: ET1702-02 IP in HeLa cell lysate Lane 3: Rabbit IgG instead of ET1702-02 in HeLa cell lysate Blocking/Dilution buffer: primary antibody dilution (K1803) Exposure time: 6 seconds; ECL: K1801 |
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