702097

antibody from Invitrogen Antibodies

Targeting: RB1 OSRC, PPP1R130, RB

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

702097

Provider

Invitrogen Antibodies

Product name

Phospho-Rb (Ser807, Ser811) Recombinant Rabbit Monoclonal Antibody (13H27L9)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with Monkey, Bovine, Pig

Antibody clone number

13H27L9

Concentration

0.5 mg/mL

References

Submitted references

Amplification of CDK4 and MDM2: a detailed study of a high-risk neuroblastoma subgroup.

Martinez-Monleon A, Kryh Öberg H, Gaarder J, Berbegall AP, Javanmardi N, Djos A, Ussowicz M, Taschner-Mandl S, Ambros IM, Øra I, Sandstedt B, Beiske K, Ladenstein R, Noguera R, Ambros PF, Gordon Murkes L, Ljungman G, Kogner P, Fransson S, Martinsson T
Scientific reports 2022 Jul 20;12(1):12420

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of COLO 205 (Lane 1, 2 & 3). The blots were probed with Anti-phospho-Rb (Ser807/Ser811) Recombinant Rabbit Monoclonal Antibody (Product # 702097, 1 µg/mL). To confirm the specificity of Phospho-Rb (Ser807/Ser811), competition was performed with the phosphopeptide (10 µg/mL) (Lane 2) and non phosphopeptide (10 µg/mL) (Lane 3). A 110 kDa band corresponding to Phospho-Rb (Ser807/Ser811) was observed as shown. The blots were detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:5000 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with wet transfer method. The membrane was probed with the relevant primary and secondary antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, COLO 205 cells were fixed and permeabilized for detection of endogenous Phospho Rb (Ser807/Ser811) using Anti- Phospho Rb (Ser807/Ser811) Recombinant Rabbit Monoclonal Antibody (Product # 702097, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of Phospho-Rb (Ser807/Ser811) protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of panels a, b and c clearly demonstrating nuclear localization of Phospho-Rb (Ser807/Ser811). Panel e) shows loss of signal by competition with the Phospho-Rb (Ser807/Ser811) phospho peptide, demonstrating antibody specificity, and panel f) demonstrates no competition with the non-phosphorylated peptide. Panel g) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.