Phospho-Bcl-2 (S70) Recombinant Rabbit Monoclonal Antibody [PSH11-02]
cat.: HA723269
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH11-02 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 26 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser70 of Human Bcl-2. |
| Positive control: | Jurrkat treated with 1μM paclitaxel overnight cell lysate, Jurkat cells treated with 1μM paclitaxel overnight. |
| Subcellular location: | Mitochondrion outer membrane, Nucleus membrane, Endoplasmic reticulum membrane, Cytoplasm. |
| Recommended Dilutions:
WB IF-Cell FC |
1:2,000 1:500 1:10,000 |
| Uniprot #: | SwissProt: P10415 Human |
| Alternative names: | Apoptosis regulator Bcl 2 Apoptosis regulator Bcl-2 Apoptosis regulator Bcl2 AW986256 B cell CLL/lymphoma 2 B cell leukemia/lymphoma 2 Bcl-2 Bcl2 BCL2_HUMAN C430015F12Rik D630044D05Rik D830018M01Rik Leukemia/lymphoma, B-cell, 2 Oncogene B-cell leukemia 2 PPP1R50 Protein phosphatase 1, regulatory subunit 50 Bcl 2 |
Images
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Fig1:
Western blot analysis of Phospho-Bcl-2 (S70) on different lysates with Rabbit anti-Phospho-Bcl-2 (S70) antibody (HA723269) at 1/2,000 dilution. Lane 1: Jurrkat cell lysate Lane 2: Jurrkat treated with 1μM paclitaxel overnight cell lysate Lane 3: Jurrkat treated with 1μM paclitaxel overnight cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 26 kDa Observed band size: 26 kDa Exposure time: 1 minute 15 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 (HA723269) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. 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 Jurkat cells untreated / treated with 1μM paclitaxel overnight labeling Phospho-Bcl-2 (S70) with Rabbit anti-Phospho-Bcl-2 (S70) antibody (HA723269) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-Phospho-Bcl-2 (S70) antibody (HA723269) 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 (HA601187, 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:
Flow cytometric analysis of untreated Jurkat cells (top) / Jurkat cells treated with 1μM paclitaxel overnight (bottom) labeling Phospho-Bcl-2 (S70). Cells were fixed and permeabilized. Then stained with the primary antibody (HA723269, 1/10,000) (right) compared with Rabbit IgG Isotype Control (1μg/mL, left). 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℃. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.