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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Flow cytometry [1]
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- Product number
- 701272 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-Rb (Ser780) Recombinant Rabbit Monoclonal Antibody (13H9L5)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with mouse, rat, Drosophila, non-human primate and rabbit based on sequence homology.
- Antibody clone number
- 13H9L5
- Concentration
- 0.5 mg/mL
Submitted references Cytoplasmic translocation of the retinoblastoma protein disrupts sarcomeric organization.
Araki K, Kawauchi K, Hirata H, Yamamoto M, Taya Y
eLife 2013 Dec 3;2:e01228
eLife 2013 Dec 3;2:e01228
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Phospho-Rb pSer780 in COLO 205 whole cell extracts using a Phospho-Rb pSer780 recombinant rabbit monoclonal antibody (Product # 701272) at a dilution of 2 µg/mL. To confirm specificity, competition was performed by preincubation with the phosphopeptide to inhibit antibody binding (lane 2). Samples were detected using chemiluminescence (ECL). Results show a band at ~110kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of RB1 (pS780) was performed by loading 30 µg of MDA-MB-231 (lane1), MCF7 (lane2), K562 (lane3), Jurkat (lane4) and COLO205 (lane5) cell lysates using Novex®NuPAGE®12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Proteins were transferred to a PVDF membrane and blocked with 5 % skim milk for 1 hour at room temperature. RB1 (pS780) was detected at ~110 kDa using RB1 (pS780) Recombinant Rabbit Monoclonal Antibody (Product # 701272) at 1-3 µg/mL in 2.5 % skim milk at 4°C overnight on a rocking platform. Goat anti-Rabbit IgG-HRP Secondary Antibody (Product # G-21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Phospho-Rb pSer780 in U2OS cells using a Phospho-Rb pSer780 recombinant rabbit monoclonal antibody (Product # 701272) 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 Rb and Image E is a composite image of cells showing inhibition of antibody binding after competition with the phosphopeptide.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of RB1 [pS780] 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 [pS780] Recombinant Rabbit Monoclonal Antibody (701272, 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.
