701059

antibody from Invitrogen Antibodies

Targeting: RB1 OSRC, PPP1R130, RB

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

701059

Provider

Invitrogen Antibodies

Product name

Anti-Phospho-Rb (Ser249, Thr252) Antibody (14H7L14), ABfinity™ Rabbit Monoclonal

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A. ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance. Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

14H7L14

Vial size

100 µg

Concentration

0.5 mg/ml

Storage

Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Phospho-Rb pSer249/pThr252 in U2OS cells using a Phospho-Rb pSer249/pThr252 recombinant rabbit monoclonal antibody (Product # 701059) followed by detection using an Alexa Fluor 488-conjugated goat anti-rabbit secondary antibody (green) (Image A). Nuclei were stained using DAPI (Image B) and actin stained with Alexa Fluor 594 phalloidin (red) (image C). Image D is a composite image showing nuclear localization of phosphorylated Rb and Image E is a composite image of cells showing competition with the phospho Rb (pSpT249/252) peptide to inhibit antibody binding.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of RB1 [pSpT249/252] was done on synchronized COLO 205 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Tritonª X-100 for 20 minutes, and blocked with 5% BSA for 1 hour at room temperature. Cells were labeled with ABfinityª RB1 [pSpT249/252] Recombinant Rabbit Monoclonal Antibody (701059, red histogram) or with rabbit isotype control (pink histogram) at 1-3 µg/million cells in 2.5% BSA. After incubation at room temperature for 2-3 hours, the cells were labeled with Alexa Fluor¨ 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune¨ Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.