Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [2]
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Validation data
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- Product number
- PA5-76788 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CPT1A Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen and the purity is > 95% (by SDS-PAGE).
- Reactivity
- Human, Rat
- Host
- Rabbit
- Isotype
- IgG
- 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 CPT1A. Samples were incubated with CPT1A polyclonal antibody (Product # PA5-76788).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Carnitine O-palmitoyl transferase 1, liver isoform was achieved by transfecting Hep G2 with Carnitine O-palmitoyl transferase 1, liver isoform specific siRNAs (Silencer® select Product # s3465, s3467). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Carnitine O-palmitoyl transferase 1, liver isoform knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with CPT1A Polyclonal Antibody (Product # PA5-76788, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:6000 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 Carnitine O-palmitoyl transferase 1, liver isoform. Additional uncharacterized band around 35kDa was also observed across the samples tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-CPT1A Polyclonal Antibody (Product # PA5-76788) and a 86kDa band corresponding to Carnitine O-palmitoyl transferase 1, liver isoform was observed across cell lines tested. Expression of CPT1A is observed to be high in MCF7 when compared to MDA-MB-231 and MCF 10A as reported in literatures. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), HEK-293 (Lane 2), SK-O-V3 (Lane 3), MCF7 (Lane 4), MDA-MB-231 (Lane 5), MCF 10A (Lane 6), PANC-1 (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were 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, 1:6000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of CPT1A in MCF7 cells. Samples were incubated with CPT1A polyclonal antibody (Product # PA5-76788).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Carnitine O-palmitoyl transferase 1, liver isoform was performed using 70% confluent log phase MCF7 and MCF 10A 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 CPT1A Polyclonal Antibody (Product # PA5-76788) 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 mitochondria like localization. Panel e represents MCF 10A cells with less expression of CPT1A. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.