WB
Western blot analysis of Phospho-Rb (S807/S811) on different lysates with Rabbit anti-Phospho-Rb (S807/S811) antibody at 1/1,000 dilution. Lane 1: K-562 cell lysate (no heat) (20 µg/Lane), Lane 2: SH-SY5Y cell lysate (no heat) (20 µg/Lane), Lane 3: K-562 cell lysate (no heat), the membrane treated with λpp for 1 hour (20 µg/Lane), Lane 4: SH-SY5Y cell lysate (no heat), the membrane treated with λpp for 1 hour (20 µg/Lane), Notice: no heat means the lysate is not boiled. Exposure time: 59 seconds; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-Phospho-Rb (S807/S811) antibody at 1/1,000 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 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.ICC/IF
Immunocytochemistry analysis of MCF7 cells labeling Phospho-Rb (S807/S811) with Rabbit anti-Phospho-Rb (S807/S811) antibody at 1/250 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-Rb (S807/S811) antibody at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of MCF7 cells labeling Phospho-Rb (S807/S811). Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
Phospho-Rb (S807/S811) was immunoprecipitated from 0.2 mg K-562 cell lysate with Rabbit anti-Phospho-Rb (S807/S811) antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-Phospho-Rb (S807/S811) antibody at 1/1,000 dilution. Mouse anti Rabbit IgG heavy chain (Fc) secondary antibody at 1/10,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input), Lane 2: Rabbit anti-Phospho-Rb (S807/S811) antibody IP in HeLa cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Phospho-Rb (S807/S811) antibody in HeLa cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 3 minutes.| Product Name | Phospho-Rb (S807/S811) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic phospho-peptide corresponding to residues surrounding Ser807/Ser811 of Human Rb. |
| Modification | P-S807/S811 |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:1000 IHC:1:1000 ICC/IF:1:250 FC:1:1000 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouse |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | RB1 |
|---|---|
| Gene Synonyms | RB pRb OSRC pp110 p105-Rb PPP1R130 p110-RB1 |
| Gene Full Name | RB transcriptional corepressor 1 |
| Gene Summary | The protein encoded by this gene is a negative regulator of the cell cycle and was the first tumor suppressor gene found. The encoded protein also stabilizes constitutive heterochromatin to maintain the overall chromatin structure. The active, hypophosphorylated form of the protein binds transcription factor E2F1. Defects in this gene are a cause of childhood cancer retinoblastoma (RB), bladder cancer, and osteogenic sarcoma. [provided by RefSeq, Jul 2008] |
| Alternative Names | Exon 17 tumor GOS561 substitution mutation causes premature stop antibody GOS563 exon 17 substitution mutation causes premature stop antibody OSRC antibody Osteosarcoma antibody p105-Rb antibody P105RB antibody PP105 antibody pp110 antibody PPP1R130 antibody pRb antibody Exon 17 tumor GOS561 substitution mutation causes premature stop antibody GOS563 exon 17 substitution mutation causes premature stop antibody OSRC antibody Osteosarcoma antibody p105-Rb antibody P105RB antibody PP105 antibody pp110 antibody PPP1R130 antibody pRb antibody Prepro retinoblastoma associated protein antibody Protein phosphatase 1 regulatory subunit 130 antibody Rb antibody RB transcriptional corepressor 1 antibody RB_HUMAN antibody RB1 antibody RB1 gene antibody Retinoblastoma 1 antibody Retinoblastoma suspectibility protein antibody Retinoblastoma-associated protein antibody |
| Molecular Weight(MW) | 106kDa(Observed band size: 110kDa) |
| Cellular Localization | Nucleus, Cytoplasm. |
WB
Western blot analysis of Phospho-Rb (S807/S811) on different lysates with Rabbit anti-Phospho-Rb (S807/S811) antibody at 1/1,000 dilution. Lane 1: K-562 cell lysate (no heat) (20 µg/Lane), Lane 2: SH-SY5Y cell lysate (no heat) (20 µg/Lane), Lane 3: K-562 cell lysate (no heat), the membrane treated with λpp for 1 hour (20 µg/Lane), Lane 4: SH-SY5Y cell lysate (no heat), the membrane treated with λpp for 1 hour (20 µg/Lane), Notice: no heat means the lysate is not boiled. Exposure time: 59 seconds; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-Phospho-Rb (S807/S811) antibody at 1/1,000 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 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.
ICC/IF
Immunocytochemistry analysis of MCF7 cells labeling Phospho-Rb (S807/S811) with Rabbit anti-Phospho-Rb (S807/S811) antibody at 1/250 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-Rb (S807/S811) antibody at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of MCF7 cells labeling Phospho-Rb (S807/S811). Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
Phospho-Rb (S807/S811) was immunoprecipitated from 0.2 mg K-562 cell lysate with Rabbit anti-Phospho-Rb (S807/S811) antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-Phospho-Rb (S807/S811) antibody at 1/1,000 dilution. Mouse anti Rabbit IgG heavy chain (Fc) secondary antibody at 1/10,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input), Lane 2: Rabbit anti-Phospho-Rb (S807/S811) antibody IP in HeLa cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Phospho-Rb (S807/S811) antibody in HeLa cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 3 minutes.| Application Notes | WB:1:1000 IHC:1:1000 ICC/IF:1:250 FC:1:1000 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | 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. |
