Scramblase 1 Recombinant Rabbit Monoclonal Antibody [JE35-30]
cat.: HA722327
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, IHC-P, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | JE35-30 |
| 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: 35 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within Human Scramblase 1 aa 11-60 / 318. |
| Positive control: | SK-MEL-28 cell lysate, HeLa cell lysate, THP-1 cell lysate, MDA-MB-231 cell lysate, SK-MEL-28, human breast cancer tissue, human spleen tissue. |
| Subcellular location: | Cell membrane, Nucleus, Membrane, Cytoplasm, perinuclear region. |
| Recommended Dilutions:
WB IF-Cell IHC-P FC IP |
1:1,000 1:100 1:200 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: O15162 Human |
| Alternative names: | Ca(2+) dependent phospholipid scramblase 1 Ca(2+)-dependent phospholipid scramblase 1 Erythrocyte phospholipid scramblase MM1 cell-derived transplantability-associated gene 1b, mouse, homolog of MmTRA1a MmTRA1b Nor1 Phospholipid scramblase 1 PL scramblase 1 PLS1_HUMAN PLSCR 1 Plscr1 Scramblase1 Tra1 Tra1a Tra1b Transplantability-associated protein 1 Tras1 Tras2 |
Images
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Fig1:
Western blot analysis of Scramblase 1 on different lysates with Rabbit anti-Scramblase 1 antibody (HA722327) at 1/1,000 dilution. Lane 1: SK-MEL-28 cell lysate Lane 2: HeLa cell lysate Lane 3: THP-1 cell lysate Lane 4: MDA-MB-231 cell lysate Lane 5: SH-SY5Y cell lysate (negative) Lysates/proteins at 20 µg/Lane. Predicted band size: 35 kDa Observed band size: 35 kDa Exposure time: 9 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 (HA722327) 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 SK-MEL-28 cells labeling Scramblase 1 with Rabbit anti-Scramblase 1 antibody (HA722327) 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-Scramblase 1 antibody (HA722327) 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 human breast cancer tissue with Rabbit anti-Scramblase 1 antibody (HA722327) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722327) 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 spleen tissue with Rabbit anti-Scramblase 1 antibody (HA722327) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722327) 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:
Scramblase 1 was immunoprecipitated from 0.2 mg SK-MEL-28 cell lysate with HA722327 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722327 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: SK-MEL-28 cell lysate (input) Lane 2: HA722327 IP in SK-MEL-28 cell lysate Lane 3: Rabbit IgG instead of HA722327 in SK-MEL-28 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 14 seconds; ECL: K1801 |
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Fig6:
Flow cytometric analysis of SK-MEL-28 cells labeling Scramblase 1. Cells were fixed and permeabilized. Then stained with the primary antibody (HA722327, 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). |
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Fig7:
Western blot analysis of Scramblase 1 on different lysates with Rabbit anti-Scramblase 1 antibody (HA722327) at 1/5,000 dilution. Lane 1: THP-1-si NT cell lysate Lane 2: THP-1-si Scramblase 1 cell lysate Lysates/proteins at 5 µg/Lane. Predicted band size: 35 kDa Observed band size: 35 kDa Exposure time: 20 seconds; ECL: K1802; 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 (HA722327) at 1/5,000 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.