Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [3]
- Immunohistochemistry [4]
- Flow cytometry [1]
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Validation data
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- Product number
- MA5-32594 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SMC3 Recombinant Rabbit Monoclonal Antibody (JM10-75)
- 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
- JM10-75
- 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 was performed using Anti-SMC3 Recombinant Rabbit Monoclonal Antibody (JM10-75) (Product # MA5-32594) and a 160 kDa band corresponding to SMC3 was observed across cell line and tissue tested. Nuclear enriched extracts (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), MCF7 (Lane 3), HCT 116 (Lane 4), U-87 MG (Lane 5), NIH/3T3 (Lane 6) and PC-12 (Lane 7), Tissue extracts of Mouse Spleen (Lane 8), Rat Spleen (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) 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). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SMC3 in different lysates using a Monoclonal antibody (Product #MA5-32594) at a dilution of 1:1,000. Positive control: Line1: HepG2, Line 2: NIH-3T3, Line 3:PC12.
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of SMC3 was achieved by transfecting HeLa with SMC3 specific siRNAs (Silencer® select Product # s17426, s17427). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the SMC3 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with SMC3 Recombinant Rabbit Monoclonal Antibody (JM10-75) (Product # MA5-32594, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to SMC3.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of SMC3 in Hela cells using a SMC3 Monoclonal antibody (Product # MA5-32594) 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 SMC3 in HepG2 cells using a SMC3 Monoclonal antibody (Product # MA5-32594) 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 SMC3 in NIH-3T3 cells using a SMC3 Monoclonal antibody (Product # MA5-32594) as seen in red. The nuclear counter stain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of SMC3 of paraffin-embedded Human tonsil tissue using a SMC3 Monoclonal antibody (Product #MA5-32594). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of SMC3 of paraffin-embedded Human colon cancer tissue using a SMC3 Monoclonal antibody (Product #MA5-32594). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of SMC3 of paraffin-embedded Mouse colon tissue using a SMC3 Monoclonal antibody (Product #MA5-32594). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of SMC3 of paraffin-embedded Zebrafish tissue using a SMC3 Monoclonal antibody (Product #MA5-32594). Counter stained with hematoxylin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometric analysis of SMC3 in Hela cells using a SMC3 Monoclonal Antibody (Product # MA5-32594) 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.