Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [4]
- Immunohistochemistry [5]
- Flow cytometry [1]
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Validation data
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- Product number
- MA5-32638 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-RNA pol II CTD (Ser5) Recombinant Rabbit Monoclonal Antibody (JM51-21)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- Recombinant rabbit monoclonal antibodies are produced using in vitro expression systems. The expression systems are developed by cloning in the specific antibody DNA sequences from immunoreactive rabbits. Then, individual clones are screened to select the best candidates for production. The advantages of using recombinant rabbit monoclonal antibodies include: better specificity and sensitivity, lot-to-lot consistency, animal origin-free formulations, and broader immunoreactivity to diverse targets due to larger rabbit immune repertoire.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- JM51-21
- Vial size
- 100 µL
- Concentration
- 1 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
- Western blot analysis of Phospho-POLR2A (Ser5) in different lysates using a Monoclonal antibody (Product #MA5-32638) at a dilution of 1:500. Positive control: Line 1: Hela, Line 2: MCF-7.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Phospho-RNA pol II CTD (Ser5) Recombinant Rabbit Monoclonal Antibody (JM51-21) (Product # MA5-32638) and a 250 kDa band corresponding to Phospho-RNA pol II CTD (Ser5) was observed across cell lines tested. Nuclear enriched extracts (40 µg lysate) of HeLa (Lane 1), HeLa treated with Human IFN Gamma (200U/mL for 24hrs) (Lane 2), Raji (Lane 3), NIH/3T3 (Lane 4), MCF7 (Lane 5) and HCT 116 (Lane 6) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were equilibrated with 20% ethanol and then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:20000 dilution using the iBright™ FL1500 Imaging System (Product # A44115). Increase in intensity was observed with Human IFN Gamma treatment.Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of Phospho-POLR2A (Ser5) in Hela cells using a Phospho-POLR2A (Ser5) Monoclonal antibody (Product # MA5-32638) as seen in red. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of Phospho-POLR2A (Ser5) in MCF-7 cells using a Phospho-POLR2A (Ser5) Monoclonal antibody (Product # MA5-32638) as seen in red. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of Phospho-POLR2A (Ser5) in PC-12 cells using a Phospho-POLR2A (Ser5) Monoclonal antibody (Product # MA5-32638) as seen in red. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Phospho-RNA pol II CTD (Ser5) Recombinant Rabbit Monoclonal Antibody (JM51-21) was performed using 70% confluent log phase HCT 116 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 Phospho-RNA pol II CTD (Ser5) Recombinant Rabbit Monoclonal Antibody (JM51-21) (Product # MA5-32638) at 1:100 dilutionin 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 dilution). 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.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of Phospho-POLR2A (Ser5) of paraffin-embedded Mouse bladder tissue using a Phospho-POLR2A-Ser5 Monoclonal antibody (Product #MA5-32638). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of Phospho-POLR2A (Ser5) of paraffin-embedded Human breast cancer tissue using a Phospho-POLR2A-Ser5 Monoclonal antibody (Product #MA5-32638). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of Phospho-POLR2A (Ser5) of paraffin-embedded Human spleen tissue using a Phospho-POLR2A-Ser5 Monoclonal antibody (Product #MA5-32638). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of Phospho-POLR2A (Ser5) of paraffin-embedded Human kidney tissue using a Phospho-POLR2A-Ser5 Monoclonal antibody (Product #MA5-32638). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of Phospho-POLR2A (Ser5) of paraffin-embedded Mouse bladder tissue using a Phospho-POLR2A-Ser5 Monoclonal antibody (Product #MA5-32638). Counter stained with hematoxylin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometric analysis of Phospho-POLR2A (Ser5) in Hela cells using a Phospho-POLR2A (Ser5) Monoclonal Antibody (Product # MA5-32638) at a dilution of 1:50, as seen in red compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.