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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [6]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
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- Product number
- PA5-29727 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- DBT Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: H1299, HepG2, mouse liver, rat liver. Predicted reactivity: Mouse (95%), Rat (95%), Zebrafish (82%), Xenopus laevis (88%), Chicken (84%), Rhesus Monkey (100%), Chimpanzee (100%), Bovine (96%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.3 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
Submitted references Mitochondrial lipoylation integrates age-associated decline in brown fat thermogenesis.
Tajima K, Ikeda K, Chang HY, Chang CH, Yoneshiro T, Oguri Y, Jun H, Wu J, Ishihama Y, Kajimura S
Nature metabolism 2019 Sep;1(9):886-898
Nature metabolism 2019 Sep;1(9):886-898
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using DBT Polyclonal Antibody (Product # PA5-29727). Sample (30 µg of whole cell lysate). Lane A: H1299. Lane B: HepG2. 10% SDS PAGE. DBT Polyclonal Antibody (Product # PA5-29727) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using DBT Polyclonal Antibody (Product # PA5-29727). Various whole cell extracts (30 µg) were separated by 10% SDS-PAGE, and the membrane was blotted with DBT Polyclonal Antibody (Product # PA5-29727) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- DBT Polyclonal Antibody detects DBT protein by western blot analysis. A. 50 µg mouse liver lysate/extract.10% SDS-PAGE. DBT Polyclonal Antibody (Product # PA5-29727) dilution: 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- DBT Polyclonal Antibody detects DBT protein by western blot analysis. A. 50 µg rat liver lysate/extract.10% SDS-PAGE. DBT Polyclonal Antibody (Product # PA5-29727) dilution: 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of DBT was achieved by transfecting LNCaP cells with DBT specific siRNAs (Silencer® select Product # s3956, s3957). Western blot analysis (Fig. a) was performed using whole cell extracts from the DBT knockdown cells (lane 2) and non-specific scrambled siRNA transfected cells (lane 1). The blots were probed with DBT Polyclonal Antibody (Product # PA5-29727, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 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 DBT.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), A-431 (Lane 2), LNCaP (Lane 3), KARPAS 299 (Lane 4), T-47D (Lane 5), NIH/3T3 (Lane 6), Jurkat (Lane 7), Mouse Liver (Lane 8), Rat Liver (Lane 9), Mouse Kidney (Lane 10) and Rat Kidney (Lane 11). The blot was probed with Anti-DBT Polyclonal Antibody (Product # PA5-29727, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 53 kDa band corresponding to DBT was observed across the cell lines and tissues tested.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry-Immunofluorescence analysis of DBT was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: DBT Polyclonal Antibody (Product # PA5-29727) diluted at 1:500. Blue: Hoechst 33342 staining. Scale bar = 10 µm.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of DBT was performed using 70% confluent log phase LNCaP cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with DBT Polyclonal Antibody (Product # PA5-29727) at 1:250 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 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 mitochondrial 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
- DBT Polyclonal Antibody detects DBT protein at mitochondria on mouse kidney by immunohistochemical analysis. Sample: Paraffin-embedded mouse kidney. DBT Polyclonal Antibody (Product # PA5-29727) diluted at 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
