Phospho-Rb (S795) Recombinant Rabbit Monoclonal Antibody [PSH06-89]
cat.: HA722764
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell |
| Clonality: | Monoclonal |
| Clone number: | PSH06-89 |
| 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: | 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: 106 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Ser795 of human Rb aa 776-825. |
| Positive control: | HeLa treated with 100ng/mL Nocodazole for 18 hours cell lysate, HeLa cells treated with 100ng/mL Nocodazole for 18 hours, U-2 OS cell lysate, U-2 OS treated with 50ng/mL Nocodazole for 8 hours cell lysate. |
| Subcellular location: | Nucleus. |
| Recommended Dilutions:
WB IF-Cell |
1:2,000 1:500 |
| Uniprot #: | SwissProt: P06400 Human |
| Alternative names: | Exon 17 tumor GOS561 substitution mutation causes premature stop GOS563 exon 17 substitution mutation causes premature stop OSRC Osteosarcoma p105-Rb P105RB PP105 pp110 PPP1R130 pRb Prepro retinoblastoma associated protein Protein phosphatase 1 regulatory subunit 130 Rb RB transcriptional corepressor 1 RB_HUMAN RB1 RB1 gene Retinoblastoma 1 Retinoblastoma suspectibility protein Retinoblastoma-associated protein |
Images
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Fig1:
Western blot analysis of Phospho-Rb (S795) on different lysates with Rabbit anti-Phospho-Rb (S795) antibody (HA722764) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 100ng/mL Nocodazole for 18 hours cell lysate Lane 3: U-2 OS cell lysate Lane 4: U-2 OS treated with 50ng/mL Nocodazole for 8 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 106 kDa Observed band size: 106 kDa Exposure time: Lane 1-4 (left): 40 seconds; Lane 1-4 (right): 1 minute; 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 (HA722764) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were 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 treated with 100ng/mL Nocodazole for 18 hours labeling Phospho-Rb (S795) with Rabbit anti-Phospho-Rb (S795) antibody (HA722764) 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-Phospho-Rb (S795) antibody (HA722764) 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:
Western blot analysis of Phospho-Rb (S795) on different lysates with Rabbit anti-Phospho-Rb (S795) antibody (HA722764) at 1/2,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 100ng/mL Nocodazole for 18 hours cell lysate Lane 3: HeLa treated with 100ng/mL Nocodazole for 18 hours cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 20 µg/Lane. Predicted band size: 106 kDa Observed band size: 106 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 (HA722764) 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. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.