PA5-29813

antibody from Invitrogen Antibodies

Targeting: HADHA GBP, LCEH, LCHAD, MTPA

Antibody data

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Validation data

HADHA protein structure - PA5-29813 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of HADHA was achieved by transfecting Hep G2 with HADHA specific siRNAs (Silencer® select Product # s6441). Western blot analysis (Fig. a) was performed using membrane enriched extracts from the HADHA knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with HADHA Polyclonal Antibody (Product # PA5-29813, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/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 HADHA.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of Hep G2 (Lane 1), HeLa (Lane 2), Jurkat (Lane 3), Raji (Lane 4), A-431 (Lane 5), tissue extracts of Mouse Heart (Lane 6), Mouse Liver (Lane 7) and Mouse Adipose (Lane 8). The blot was probed with Anti-HADHA Polyclonal Antibody (Product # PA5-29813, 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 75 kDa band corresponding to HADHA was observed across the cell lines and tissues tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using HADHA Polyclonal Antibody (Product # PA5-29813). Sample (30 µg of whole cell lysate). Lane A: 293T. Lane B: A431 . 7.5% SDS PAGE. HADHA Polyclonal Antibody (Product # PA5-29813) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using HADHA Polyclonal Antibody (Product # PA5-29813). Sample (50 µg of whole cell lysate). Lane A: mouse liver . 7.5% SDS PAGE. HADHA Polyclonal Antibody (Product # PA5-29813) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: HADHA Polyclonal Antibody detects HADHA protein by Western blot analysis. A.30 µg PC-12 whole cell lysate/extract.7.5 % SDS-PAGE. HADHA Polyclonal Antibody (Product # PA5-29813) dilution: 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of HADHA 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 1% BSA for 1 hour at room temperature. The cells were labeled with HADHA Polyclonal Antibody (Product # PA5-29813) at 1:100 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 SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Mitochondria (Panel c: red) was stained with Mitotracker Red CMXRos (Product # M7512). 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)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of HADHA was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: HADHA Polyclonal Antibody (Product # PA5-29813) diluted at 1:500. Red: alpha Tubulin, a cytoskeleton marker. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: HADHA Polyclonal Antibody detects HADHA protein at Mitochondia by immunofluorescent analysis. Sample: HepG2 cells were fixed in -20ºC 100% MeOH for 5 min. Green: HADHA protein stained by HADHA Polyclonal Antibody (Product # PA5-29813) diluted at 1:500. Blue: Hoechst 33343 staining.