Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [4]
- Immunohistochemistry [6]
- Flow cytometry [1]
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Validation data
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- Product number
- MA5-32598 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SDHA Recombinant Rabbit Monoclonal Antibody (JM10-83)
- 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, Zebrafish
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- JM10-83
- 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 SDHA in Jurkat cell lysates using a SDHA Monoclonal antibody (Product # MA5-32598) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of SDHA was achieved by transfecting SH-SY5Y with SDHA specific siRNAs (Silencer® select Product # S12650, S12651). Western blot analysis (Fig. a) was performed using Membrane enriched extracts from the SDHA knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with SDHA Recombinant Rabbit Monoclonal Antibody (JM10-83) (Product # MA5-32598, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 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 SDHA.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-SDHA Recombinant Rabbit Monoclonal Antibody (JM10-83)(Product # MA5-32598) and a 70kDa band corresponding to SDHA was observed across cell lines and tissues tested. Membrane enriched extracts (30 µg lysate) of SH-SY5Y (Lane 1), HeLa (Lane 2), LNCaP (Lane 3), SK-O-V3 (Lane 4) and Hep G2 (Lane 5); and tissue extracts of Mouse Kidney (Lane 6) and Rat Heart (Lane 7) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). 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, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of SDHA in MCF-7 cells using a SDHA Monoclonal antibody (Product # MA5-32598) 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)
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- Experimental details
- Immunocytochemical analysis of SDHA in HepG2 cells using a SDHA Monoclonal antibody (Product # MA5-32598) 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 SDHA in NIH-3T3 cells using a SDHA Monoclonal antibody (Product # MA5-32598) 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 SDHA was performed using 70 confluent log phase HeLa 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 SDHA Recombinant Rabbit Monoclonal Antibody (JM10-83) (Product # MA5-32598) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (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 cytoplasmic (mitochondria-like) localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemical analysis of SDHA of paraffin-embedded Zebrafish tissue using a SDHA Monoclonal antibody (Product #MA5-32598). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemical analysis of SDHA of paraffin-embedded Human liver tissue using a SDHA Monoclonal antibody (Product #MA5-32598). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of SDHA of paraffin-embedded Human kidney tissue using a SDHA Monoclonal antibody (Product #MA5-32598). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of SDHA of paraffin-embedded Mouse testis tissue using a SDHA Monoclonal antibody (Product #MA5-32598). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunohistochemical analysis of SDHA of paraffin-embedded Mouse skeletal muscle tissue using a SDHA Monoclonal antibody (Product #MA5-32598). Counter stained with hematoxylin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of SDHA of paraffin-embedded Mouse colon tissue using a SDHA Monoclonal antibody (Product #MA5-32598). Counter stained with hematoxylin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow Cytometric analysis of SDHA in Hela cells using a SDHA Monoclonal Antibody (Product # MA5-32598) 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.