Phospho-PPP1CA (T320) Recombinant Rabbit Monoclonal Antibody [PSH10-27]
cat.: HA723194
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH10-27 |
| 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: 38 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Thr320 of Human PPP1CA. |
| Positive control: | HeLa treated with 100nM Calyculin A for 30 minutes cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate, human breast cancer tissue, human testis tissue, rat testis tissue. |
| Subcellular location: | Cytoplasm, Nucleus, nucleoplasm, nucleolus. |
| Recommended Dilutions:
WB IHC-P IP |
1:2,000 1:20,000 1-2μg/sample |
| Uniprot #: | SwissProt: P62136 Human | P62137 Mouse | P62138 Rat |
| Alternative names: | Alpha isoform serine threonine protein phosphatase PP1alpha 1 catalytic subunit Catalytic subunit EC 3.1.3.16 MGC15877 MGC1674 PP 1A PP-1A PP1A PP1A_HUMAN PP1alpha PP2C ALPHA PP2CA Ppp1ca Protein Phosphatase 2C Alpha Isoform Serine threonine protein phosphatase PP1 alpha catalytic subunit Serine threonine protein phosphatase PP1 alpha catalytic subunit protein phosphatase 1 Serine/threonine-protein phosphatase PP1-alpha catalytic subunit |
Images
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Fig1:
Western blot analysis of Phospho-PPP1CA (T320) on different lysates with Rabbit anti-Phospho-PPP1CA (T320) antibody (HA723194) at 1/2,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate Lane 3: NIH/3T3 cell lysate Lane 4: PC-12 cell lysate Lane 5: PC-12 treated with 100ng/mL Nocodazole for 18 hours cell lysate Lane 6: HeLa treated with 100nM Calyculin A for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 38 kDa Observed band size: 35 kDa Exposure time: 3 minutes; 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 (HA723194) at 1/2,000 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:
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Phospho-PPP1CA (T320) antibody (HA723194) at 1/20,000 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 (HA723194) at 1/20,000 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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Fig3:
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-Phospho-PPP1CA (T320) antibody (HA723194) at 1/20,000 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 (HA723194) at 1/20,000 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-Phospho-PPP1CA (T320) antibody (HA723194) at 1/20,000 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 (HA723194) at 1/20,000 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:
Phospho-PPP1CA (T320) was immunoprecipitated from 0.2 mg HeLa treated with 50mM Calyculin A for 3 hours cell lysate with HA723194 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723194 at 1/2,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa treated with 50mM Calyculin A for 3 hours cell lysate (input) Lane 2: HA723194 IP in HeLa treated with 50mM Calyculin A for 3 hours cell lysate Lane 3: Rabbit IgG instead of HA723194 in HeLa treated with 50mM Calyculin A for 3 hours cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 1 minute 23 seconds; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.