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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [9]
- Immunocytochemistry [2]
- Immunohistochemistry [4]
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- Product number
- MA5-17264 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Citrate Synthase Monoclonal Antibody (GT1761)
- Antibody type
- Monoclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, HeLa, HepG2, A375, mouse brain, rat brain, zebrafish brain, zebrafish heart.
- Antibody clone number
- GT1761
- Concentration
- 1 mg/mL
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot analysis of Citrate synthase using A) 30 µg 293T whole cell lysate (B) 30 µg A431 whole cell lysate (C) 30 µg HeLa whole cell lysate (D) 30 µg HepG2 whole cell lysate and E) 30 µg A375 whole cell lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a Citrate synthase monoclonal antibody (Product # MA5-17264) at a dilution of 1:1000.
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed using Anti-Citrate Synthase Monoclonal Antibody (GT1761) (Product # MA5-17264) and a 51 kDa band corresponding to Citrate Synthase was observed across the cell lines tested and tissues tested. Whole cell extracts (30 µg lysate) of LNCaP (Lane 1), HeLa (Lane 2), Hep G2 (Lane 3), MCF7 (Lane 4), Mouse Brain (Lane 5), Rat Kidney (Lane 6), Rat Brain (Lane 7), Mouse Liver (Lane 8) 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 mg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:2000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Citrate Synthase was performed by separating 50 µg of rat tissue extract by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a Citrate Synthase Monoclonal Antibody (GT1761) (Product # MA5-17264) at a dilution of 1:1000. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using Citrate Synthase Monoclonal Antibody (GT1761) (Product # MA5-17264). HepG2 and mitochondria extracts (30 µg) were separated by SDS-PAGE, and the membrane was blotted with Citrate synthetase antibody [GT1761] Citrate Synthase Monoclonal Antibody (GT1761) (Product # MA5-17264) diluted at 1:1,000. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Citrate synthetase antibody [GT1761] detects CS protein by western blot analysis. A. 30 µg zebrafish brain lysate/extract.10% SDS-PAGE. Citrate synthetase antibody [GT1761] Citrate Synthase Monoclonal Antibody (GT1761) (Product # MA5-17264) dilution: 1:1,000. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Citrate synthetase antibody [GT1761] detects CS protein by western blot analysis. A. 30 µg zebrafish heart lysate/extract.10% SDS-PAGE. Citrate synthetase antibody [GT1761] Citrate Synthase Monoclonal Antibody (GT1761) (Product # MA5-17264) dilution: 1:1,000. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Citrate synthetase antibody [GT1761] detects CS protein by western blot analysis. A. 50 µg mouse brain lysate/extract.12% SDS-PAGE. Citrate synthetase antibody [GT1761] Citrate Synthase Monoclonal Antibody (GT1761) (Product # MA5-17264) dilution: 1:1,000. The HRP-conjugated anti-mouse IgG antibody was used to detect the primary antibody.
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Citrate Synthase was achieved by transfecting HepG2 cells with Citrate Synthase specific siRNAs (Silencer® select Product # S3582, S3583). Western Blot analysis (Fig. a) was performed using Whole cell extracts from the Citrate Synthase knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The Blot was probed with Citrate Synthase Monoclonal Antibody (GT1761) (Product # MA5-17264, 1:1000 ) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000). 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 Citrate Synthase.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed using Anti-Citrate Synthase Monoclonal Antibody (GT1761) (Product # MA5-17264) and a 51 kDa band corresponding to Citrate Synthase was observed across the cell lines tested and tissues tested. Whole cell extracts (30 µg lysate) of LNCaP (Lane 1), HeLa (Lane 2), Hep G2 (Lane 3), MCF7 (Lane 4), Mouse Brain (Lane 5), Rat Kidney (Lane 6), Rat Brain (Lane 7), Mouse Liver (Lane 8) 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 mg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:2000) 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
- Immunocytochemistry-Immunofluorescence analysis of Citrate Synthase was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: Citrate Synthase Monoclonal Antibody (GT1761) (Product # MA5 17264) diluted at 1:200. Red: phalloidin, a cytoskeleton marker. Blue: Hoechst 33342 staining.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Citrate Synthase was performed using 70% confluent log phase Hep G2 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 Citrate Synthase Monoclonal Antibody (GT1761) (Product # MA5-17264) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723), (1:2000), 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 Mitochondria 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
- Citrate synthetase antibody [GT1761] detects Citrate synthetase protein at mitochondria on mouse heart by immunohistochemical analysis. Sample: Paraffin-embedded mouse heart. Citrate synthetase antibody [GT1761] (Product # MA5-17264) dilution: 1:200. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Citrate synthetase antibody [GT1761] detects Citrate synthetase protein at mitochondria on mouse heart by immunohistochemical analysis. Sample: Paraffin-embedded mouse heart. Citrate synthetase antibody [GT1761] (Product # MA5-17264) dilution: 1:200. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Citrate synthetase antibody [GT1761] detects Citrate synthetase protein at mitochondria on mouse kidney by immunohistochemical analysis. Sample: Paraffin-embedded mouse kidney. Citrate synthetase antibody [GT1761] (Product # MA5-17264) dilution: 1:200. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Citrate synthetase antibody detects CS protein at cytosol on H1299 xenograft by immunohistochemical analysis. Sample: Paraffin-embedded H1299 xenograft. Citrate synthetase antibody (Product # MA5-17264) dilution: 1:200. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
