- PA5-29956 - Invitrogen Antibodies ACAA1 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of ACAA1 was achieved by transfecting Hep G2 with ACAA1 specific siRNAs (Silencer® select Product # s879, s881). Western blot analysis (Fig. a) was performed using whole cell extracts from the ACAA1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with ACAA1 Polyclonal Antibody (Product # PA5-29956, 1:2000 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 ACAA1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), A549 (Lane 2), HeLa (Lane 3), SK-BR-3 (Lane 4), MCF7 (Lane 5), RAW 264.7 (Lane 6), Jurkat (Lane 7), tissue extract of Mouse Liver (Lane 8), Rat Liver (Lane 9) and Mouse Kidney (Lane 10). The blot was probed with Anti-ACAA1 Polyclonal Antibody (Product # PA5-29956, 1:2000 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 44 kDa band corresponding to ACAA1 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 ACAA1 Polyclonal Antibody (Product # PA5-29956). Sample (30 µg of whole cell lysate). Lane A: A549. Lane B: Hela. 10% SDS PAGE. ACAA1 Polyclonal Antibody (Product # PA5-29956) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using ACAA1 Polyclonal Antibody (Product # PA5-29956). Sample (50 µg of whole cell lysate). Lane A: mouse Liver. 10% SDS PAGE. ACAA1 Polyclonal Antibody (Product # PA5-29956) diluted at 1:1,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of ACAA1 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 ACAA1 Polyclonal Antibody (Product # PA5-29956) 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). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing punctate cytoplasmic 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 ACAA1 was performed in HeLa cells fixed in ice-cold MeOH for 5 min. Green: ACAA1 Polyclonal Antibody (Product # PA5-29956) diluted at 1:250. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: ACAA1 Polyclonal Antibody detects ACAA1 protein at cytosol on mouse stomach by immunohistochemical analysis. Sample: Paraffin-embedded mouse stomach. ACAA1 Polyclonal Antibody (Product # PA5-29956) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

PA5-29956