Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Immunocytochemistry [6]
- Immunoprecipitation [1]
- Immunohistochemistry [2]
- Other assay [1]
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Validation data
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- Product number
- PA5-27215 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Rb Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: Jurkat, A431, CL1-0, CL1-5. Predicted reactivity: Mouse (96%), Rat (96%), Pig (96%), Chicken (81%), Rhesus Monkey (100%), Bovine (97%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1.47 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry-Immunofluorescence analysis of Rb was performed in MCF7 cells fixed in 4% paraformaldehyde at RT for 15 min. Green: Rb Polyclonal Antibody (Product # PA5-27215) diluted at 1:500. Blue: Hoechst 33342 staining. Scale bar = 10 µm.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Rb Polyclonal Antibody detects Rb protein at nucleus by immunofluorescent analysis. Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: Rb stained by Rb Polyclonal Antibody (Product # PA5-27215) diluted at 1:500. Red: phalloidin, a cytoskeleton marker, diluted at 1:200. Scale bar= 10 µm.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Retinoblastoma-associated protein was performed using 70% confluent log phase MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Rb Polyclonal Antibody (Product # PA5-27215) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry-Immunofluorescence analysis of Rb was performed in MCF7 cells fixed in 4% paraformaldehyde at RT for 15 min. Green: Rb Polyclonal Antibody (Product # PA5-27215) diluted at 1:500. Blue: Hoechst 33342 staining. Scale bar = 10 µm.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Retinoblastoma-associated protein was performed using 70% confluent log phase MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Rb Polyclonal Antibody (Product # PA5-27215) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Rb Polyclonal Antibody detects Rb protein at nucleus by immunofluorescent analysis. Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: Rb stained by Rb Polyclonal Antibody (Product # PA5-27215) diluted at 1:500. Red: phalloidin, a cytoskeleton marker, diluted at 1:200. Scale bar= 10 µm.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Rb Polyclonal Antibody immunoprecipitates retinoblastoma 1 protein in IP experiments. IP samples: HeLa whole cell extract. A. Control with 4 µg of preimmune Rabbit IgG. B. Immunoprecipitation of retinoblastoma 1 protein by 4 µg Rb Polyclonal Antibody (Product # PA5-27215). 7.5 % SDS-PAGE. The immunoprecipitated retinoblastoma 1 protein was detected by Rb Polyclonal Antibody (Product # PA5-27215) diluted at 1:500.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Rb Polyclonal Antibody detects Rb protein at nucleus on human colon carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human colon carcinoma. Rb Polyclonal Antibody (Product # PA5-27215) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Rb Polyclonal Antibody detects Rb protein at nucleus by immunohistochemical analysis. Sample: Paraffin-embedded mouse eye. Green: Rb stained by Rb Polyclonal Antibody (Product # PA5-27215) diluted at 1:500. Red: beta Tubulin 3/ Tuj1, a cytoskeleton marker, stained by beta Tubulin 3/ Tuj1 antibody [GT11710] diluted at 1:500. Blue: Fluoroshield with DAPI. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Rb Polyclonal Antibody immunoprecipitates retinoblastoma 1 protein in IP experiments. IP samples: HeLa whole cell extract. A. Control with 4 µg of preimmune Rabbit IgG. B. Immunoprecipitation of retinoblastoma 1 protein by 4 µg Rb Polyclonal Antibody (Product # PA5-27215). 7.5 % SDS-PAGE. The immunoprecipitated retinoblastoma 1 protein was detected by Rb Polyclonal Antibody (Product # PA5-27215) diluted at 1:500.