- PA5-31157 - Invitrogen Antibodies HADH antibody

Chazarin B, Benhaim-Delarbre M, Brun C, Anzeraey A, Bertile F, Terrien J
International journal of molecular sciences 2022 Apr 12;23(8)

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of HADH using A) 30 µg 293T whole cell lysate (B) 30 µg A431 whole cell lysate (C) 30 µg HeLa whole cell lysate and D) 30 µg HepG2 whole cell lysate. Samples were loaded onto a 10% SDS-PAGE gel and probed with a HADH polyclonal antibody (Product # PA5-31157) at a dilution of 1:1000.

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), HeLa (Lane 2), A-431 (Lane 3), MSC (Lane 4), MSC differentiated to adipocytes (Lane 5) and tissue extract of Mouse Liver (Lane 6). The blot was probed with Anti-HADH Polyclonal Antibody (Product # PA5-31157, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 ug/ml, 1:4000 dilution). A 34 kDa band corresponding to HADH was observed across cell lines and tissue tested and was also enhanced in MSC after differentiating it to adipocytes.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HADH was performed by separating 50 µg of rat tissue extract by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a HADH Polyclonal Antibody (Product # PA5-31157) at a dilution of 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using HADH Polyclonal Antibody (Product # PA5-31157). Sample (50 µg of whole cell lysate). Lane A: mouse liver. 10% SDS PAGE. HADH Polyclonal Antibody (Product # PA5-31157) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using HADH Polyclonal Antibody (Product # PA5-31157). Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with HADH Polyclonal Antibody (Product # PA5-31157) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HADH was performed by separating 50 µg of rat tissue extract by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a HADH Polyclonal Antibody (Product # PA5-31157) at a dilution of 1:1000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of HADH was performed using 70% confluent log phase HeLa 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 HADH Polyclonal Antibody (Product # PA5-31157) 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 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: HADH Polyclonal Antibody detects HADH protein at Mitochondria by immunofluorescent analysis. Sample: HeLa cells were fixed in 2% paraformaldehyde/culture medium at 37ºC for 30 min. Green: HADH protein stained by HADH Polyclonal Antibody (Product # PA5-31157) diluted at 1:500. Blue: Hoechst 33343 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: HADH Polyclonal Antibody detects HADH protein at mitochondria by immunofluorescent analysis. Sample: HepG2 cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: HADH stained by HADH Polyclonal Antibody (Product # PA5-31157) diluted at 1:500. Red: alpha Tubulin, a cytoskeleton marker, stained by alpha Tubulin Polyclonal Antibody [GT114] (Product # MA5-31466) diluted at 1:1,000. Blue: Fluoroshield with DAPI .

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: HADH Polyclonal Antibody detects HADH protein at mitochondria on human colon carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded colon carcinoma. HADH Polyclonal Antibody (Product # PA5-31157) dilution: 1:250. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Regulation of liver lipid metabolism in grey mouse lemurs during winter. The relative abundance of liver proteins involved in carbohydrate metabolism in early (EW) and late (LW) winter mouse lemurs (N = 5/group) is shown using mean fold changes in LW vs EW (panel A ; presence of several isoforms of the same protein are indicated with asterisks (*). Food efficiency (FE), calculated as the ratio between food intake (in Kcal/day) and BMC were significantly different between EW and LW animals (panel B ), and the respiratory exchange ratio measured in EW (black line) vs. LW (grey line) reflected the main energy substrates used, depending on time-of-day (panel C ). Protein expression levels of mitochondrial hydroxyacyl-coenzyme A dehydrogenase (HADH), citrate synthase (CS) and uncoupling protein 3 (UCP3) were measured using Western-blot analysis in liver samples from mouse lemurs in EW and LW (blots shown in panel D and relative expression in (panel E ); N = 4-5/group; results are presented as the means +- SEM). XBP1 gene expression levels did not differ in the liver of adult male mouse lemurs between EW and LW (panel F ). * and ** indicate significance at p < 0.05 and 0.01 between EW and LW, respectively.

PA5-31157