重组Anti-Rb (phospho S807)抗体[EPR17732]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Rb (phospho S807) antibody [EPR17732] (AB184796)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% tritonX-100 permeabilized MCF7 (human breast adenocarcinoma cell line) cells, Serum starved and non-starved, labeling Rb (phospho S807) with ab184796 at 1/200 dilution followed by Goat anti-Rabbit secondary IgG AlexaFluor®488 (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing positive staining on MCF7 cells. The number of positive cells increased after treatment with FBS (fetal bovine serum) for 48 hours, then decreased after Lambda Protein Phosphatase treatment (31℃ for 2 hours ).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S807) antibody [EPR17732] (AB184796)

Immunohistochemical analysis of paraffin-embedded human Squamous cells carcinoma of lung tissue labeling Rb (phospho S807) with ab184796 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on squamous cells carcinoma of lung is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S807) antibody [EPR17732] (AB184796)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Rb (phospho S807) with ab184796 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on epithelial cells of Human colon is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S807) antibody [EPR17732] (AB184796)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling Rb (phospho S807) with ab184796 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on lymphocytes of rat spleen is observed. Counter stained with Hematoxylin.

Negative control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Rb (phospho S807) antibody [EPR17732] (AB184796)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Rb (phospho S807) with ab184796 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on lymphocytes of mouse spleen is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-Rb (phospho S807) antibody [EPR17732] (AB184796)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Rb (phospho S807) antibody [EPR17732] (ab184796) at 1/1000 dilution

All lanes:

MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 106 kDa

Observed band size: 106 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-Rb (phospho S807) antibody [EPR17732] (AB184796)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Rb (phospho S807) antibody [EPR17732] (ab184796) at 1/20000 dilution

Lane 1:

Jurkat (human T cell leukemia cell line from peripheral blood) serum starved for 44 h. Whole cell lysates at 15 µg

Lane 2:

Jurkat (human T cell leukemia cell line from peripheral blood) serum starved for 24 h. Then 10% FBS incubated for 20 h. Whole cell lysates at 15 µg

Lane 3:

Jurkat (human T cell leukemia cell line from peripheral blood) serum starved for 24 h. Then 10% FBS incubated for 20 h. Whole cell lysates. Then the membrane was incubated with phosphatase. at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 106 kDa

Observed band size: 106 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-Rb (phospho S807) antibody [EPR17732] (AB184796)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Rb (phospho S807) antibody [EPR17732] (ab184796) at 1/5000 dilution

Lane 1:

K562 (human chronic myelogenous leukemia cells from bone marrow) whole cell lysates at 10 µg

Lane 2:

K562 (human chronic myelogenous leukemia cells from bone marrow) whole cell lysates treated with Lambda Phosphatase at 10 µg

Secondary

Lane 1:

Goat Anti-Rabbit IgG (H+L), Peroxidase conjugated at 1/1000 dilution

Lane 2:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 106 kDa

Observed band size: 106 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-Rb (phospho S807) antibody [EPR17732] (AB184796)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Rb (phospho S807) antibody [EPR17732] (ab184796) at 1/20000 dilution

Lane 1:

K562 (human chronic myelogenous leukemia cells from bone marrow) whole cell lysates at 10 µg

Lane 2:

K562 (human chronic myelogenous leukemia cells from bone marrow) whole cell lysates serum starved for 2 days at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 106 kDa

Observed band size: 106 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Rb (phospho S807) antibody [EPR17732] (AB184796)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Rb (phospho S807) antibody [EPR17732] (ab184796) at 1/2000 dilution

All lanes:

C6 (rat glial tumor cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 106 kDa

Observed band size: 106 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-Rb (phospho S807) antibody [EPR17732] (AB184796)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Rb (phospho S807) antibody [EPR17732] (ab184796) at 1/2000 dilution

All lanes:

Mouse embryo lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 106 kDa

Observed band size: 106 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Rb (phospho S807) antibody [EPR17732] (AB184796)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Rb (phospho S807) antibody [EPR17732] (ab184796) at 1/1000 dilution

All lanes:

L-929 (mouse connective tissue fibroblast cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 106 kDa

Observed band size: 106 kDa

false

Exposure time: 3min

• Dot

Supplier Data

Dot Blot - Anti-Rb (phospho S807) antibody [EPR17732] (AB184796)

Dot blot analysis of Rb (phospho S807) peptide (Lane 1), and non-phospho peptide (Lane 2), labeled using ab184796 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution.

Blocking/Dilution buffer : 5% NFDM/TBST.

• WB

CiteAb

Western blot - Anti-Rb (phospho S807) antibody [EPR17732] (AB184796)

Rb (phospho S807) western blot using anti-Rb (phospho S807) antibody [EPR17732] ab184796. Publication image and figure legend from Bouclier, C., Simon, M., et al., 2020, Theranostics, PubMed 32104498.

ab184796 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab184796 please see the product overview.

P2short inhibits CDK4 activity and NSCLC proliferation but does not block cell cycle arrest. A. p107Rb phosphorylation (Ser807/811) was determined by Western blot of A549 cell extracts prepared following treatment with Abemaciclib or P2short peptide. Actin levels were detected to control constant protein loading. Left panel : Rb phosphorylation after 24 and 48 h, following treatment with 20 μM Abemaciclib or P2short peptide. Right panel : quantification of the relative Rb phosphorylation/actin levels in after 24 h following treatment with 20 μM Ctrl stapled peptide or different μM concentrations of P2short peptide. B-E.Proliferation assays.B. A549 cells were treated with different concentrations of Ctrl stapled peptide, P2, P2shortA and Abemaciclib. C. A549 cells were treated with combined administrations of P2shortA peptide (10 μM) and Abemaciclib (500 nM) Bars are averages of three independent experiments (n=4/experiment) ± SD. Means were compared with t test, * p<0.05,** p<0.02, *** p<0.01. D-E. PC9, H358 and A549 cells were treated with different concentrations of P2shortA (D) or Abemaciclib (E). Results are presented as average of at least two independent experiments (n = 4/experiment) ± standard deviation. F. Flow cytometry of A549 cells treated with DMSO, 0.5 or 5 μM Abemaciclib (Abe) and/or 1 or 10 μM Ctrl stapled or P2shortA peptide for 24 h. Means were compared with t test, * p<0.2,** p<0.1, *** p<0.05.

false