SAGE1 Recombinant Rabbit Monoclonal Antibody [PSH05-89]
cat.: HA751036
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IHC-P, IF-Cell, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH05-89 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4). |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 99 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within Human SAGE1 aa 75-733. |
| Positive control: | K-562 cell lysates, K-562, human testis tissue. |
| Subcellular location: | Nuclear body. Nucleoplasm. |
| Recommended Dilutions:
WB IHC-P IF-Cell IP |
1:1,000 1:1,000 1:500 1-2μg/sample |
| Uniprot #: | SwissProt: Q9NXZ1 Human |
| Alternative names: | Cancer/testis antigen 14 CT14 RP11-97N5.1 SAGE SAGE1 SAGE1_HUMAN Sarcoma antigen 1 |
Images
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Fig1:
Western blot analysis of SAGE1 on K-562 cell lysates with Rabbit anti-SAGE1 antibody (HA751036) at 1/1,000 dilution. Lysates/proteins at 20 µg/Lane. Predicted band size: 99 kDa Observed band size: 120 kDa Exposure time: 20 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 (HA751036) 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 K-562 cells labeling SAGE1 with Rabbit anti-SAGE1 antibody (HA751036) at 1/500 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-SAGE1 antibody (HA751036) at 1/500 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 human testis tissue with Rabbit anti-SAGE1 antibody (HA751036) 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 (HA751036) 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:
SAGE1 was immunoprecipitated from 0.2 mg K-562 cell lysate with HA751036 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751036 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: K-562 cell lysate (input) Lane 2: HA751036 IP in K-562 cell lysate Lane 3: Rabbit IgG instead of HA751036 in K-562 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 3 minutes; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.