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9306
PhosphoPlus® Rb (Ser780, Ser807/811) Antibody Kit
Primary Antibodies
PhosphoPlus Antibody Kit

PhosphoPlus® Rb (Ser780, Ser807/811) Antibody Kit #9306

Citations (1)
Confocal immunofluorescent analysis of fixed frozen mouse small intestine labeled with Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb (left, green) and co-labeled with DyLight 554 Phalloidin #13054 (right, red) and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
Confocal immunofluorescent analysis of fixed frozen mouse testis labeled with Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb (left, green) and co-labeled with DyLight 554 Phalloidin #13054 (right, red) and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
Confocal immunofluorescent analysis of fixed frozen mouse thymus labeled with Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb (left, green) and co-labeled with DyLight 554 Phalloidin #13054 (right, red) and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
Simple Western™ analysis of lysates (0.1 mg/mL) from MCF-7 cells using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb #8516. The virtual lane view (left) shows the target band (as indicated) at 1:10 dilution of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 dilution of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from WI-38 cells, serum-starved for 3 days (-) or serum-starved for 3 days followed by treatment with 10% serum for 2 days (+), using Phospho-Rb (Ser780) (D59B7) Rabbit mAb.
Western blot analysis of extracts from MCF7 cells, untreated (-) or treated with calf intestinal phosphatase (CIP) and λ phosphatase (+), using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb (upper) or Rb (4H1) Mouse mAb #9309 (lower).
Western blot analysis of Rb-C Fusion Protein #6022 (amino acids 701-928 of Rb fused to MBP) before (-) and after (+) in vitro phosphorylation by cdc2/cyclin B Protein Kinase (New England Biolabs #P6020), using Phospho-Rb (Ser795) Antibody #9301 (left) or the control antibody (right).
Western blot analysis of extracts from HeLa cells (lane 1) or Rb knock-out cells (lane 2) using Rb (4H1) Mouse mAb #9309 (upper), and β-actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the Rb knock-out HeLa cells confirms specificity of the antibody for Rb.
Western blot analysis of phosphorylated or nonphosphorylated recombinant, truncated Rb, without or with Rb blocking peptide, using Phospho-Rb (Ser780) (D59B7) Rabbit mAb.
Western blot analysis of extracts from WI-38 cells, serum-starved for 3 days (-) or serum-starved for 3 days followed by treatment with 10% serum for 2 days (+), using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb.
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-), SignalSilence® Rb siRNA I #6451 (+) or SignalSilence® Rb siRNA II (+), using Rb (4H1) Mouse mAb #9309 and α-Tubulin (11H10) Rabbit mAb #2125. The Rb (4H1) Mouse mAb confirms silencing of Rb expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of Rb siRNA.
Immunoprecipitation of phospho-Rb (Ser780) from WI-38 cell extracts using Phospho-Rb (Ser780) (D59B7) Rabbit mAb (lane 2). Western blot was performed using the same antibody. Lane 1 is 10% input.
Immunoprecipitation of phospho-Rb (Ser807/811) from Cos cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb.
Western blot analysis of extracts from COS-7 cells, untreated or hydroxyurea-treated (G1/S), using Rb (4H1) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing nuclear localization, using Rb (4H1) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Rb (4H1) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, control (left) or λ phosphatase-treated (right), using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human esophageal carcinoma using Rb (4H1) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded mouse spleen using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human T cell lymphoblastic lymphoma using Rb (4H1) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded human ovarian serous adenocarcinoma using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded human gastric adenocarcinoma using Rb (4H1) Mouse mAb.
Confocal immunofluorescent analysis of MCF7 (left) and BT-549 (right) cells, untreated (upper) or λ phosphatase-treated (lower) using Phospho-Rb (Ser807/Ser811) (D20B12) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Confocal immunofluorescent image of SH-SY5Y cells, using RB (4H1) Mouse mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
Flow cytometric analysis of Jurkat cells using Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb and Propidium Iodide (PI/RNase Staining Solution #4087 to measure DNA content. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Flow cytometric analysis of Jurkat cells, using Rb (4H1) Mouse mAb versus propidium iodide (DNA content). The box indicates Rb positive cells.
Chromatin immunoprecipitations were performed with cross-linked chromatin from Raji cells and either Rb (4H1) Mouse mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using using SimpleChIP® Human Timeless Intron 1 Primers #7001, human DHFR promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Inquiry Info.# 9306

Product Description

The PhosphoPlus® Rb (Ser780, Ser807/811) Antibody Kit provides reagents and protocols to investigate cell cycle progression within cells. The kit contains a total Rb antibody, two distinct phospho-Rb antibodies, and Rb control proteins along with secondary antibodies and reagents to perform up to 10 western blots.

Specificity / Sensitivity

Phospho-Rb (Ser780) (D59B7) Rabbit mAb and Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb detect endogenous levels of Rb only when phosphorylated at Ser780 and Ser807/811, respectively. Rb (4H1) Mouse mAb detects endogenous levels of total Rb protein. The antibody does not cross-react with the Rb homologues p107 or p130, or with other proteins.

Source / Purification

Phospho-Rb (Ser780) (D59B7) Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser780 of human Rb protein. Phospho-Rb (Ser807/811) (D20B12) XP® Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser807/811 of human Rb protein. Rb (4H1) monoclonal antibody is produced by immunizing animals with Rb-C Fusion Protein #6022, which contains residues 701-928 of human Rb .

Background

The retinoblastoma tumor suppressor protein Rb regulates cell proliferation by controlling progression through the restriction point within the G1-phase of the cell cycle (1). Rb has three functionally distinct binding domains and interacts with critical regulatory proteins including the E2F family of transcription factors, c-Abl tyrosine kinase, and proteins with a conserved LXCXE motif (2-4). Cell cycle-dependent phosphorylation by a CDK inhibits Rb target binding and allows cell cycle progression (5). Rb inactivation and subsequent cell cycle progression likely requires an initial phosphorylation by cyclin D-CDK4/6 followed by cyclin E-CDK2 phosphorylation (6). Specificity of different CDK/cyclin complexes has been observed in vitro (6-8) and cyclin D1 is required for Ser780 phosphorylation in vivo (9).

Pathways

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Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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