- 43-9800 - Invitrogen Antibodies ATP5F1A antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), NIH/3T3 (Lane 2), Hep G2 (Lane 3), COS-7 (Lane 4), MCF7 (Lane 5), MDA-MB-231 (Lane 6), RAW 264.7 (Lane 7), tissue extracts of Mouse Brain (Lane 8), Rat Brain (Lane 9), Mouse Colon (Lane 10) and Mouse lung (Lane 11). The blot was probed with Anti-ATP5A1 Monoclonal Antibody (15H4C4) (Product # 43-9800, 1 µg/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). A 55 kDa band corresponding to ATP5A1 was detected across the cell lines and tissues tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ATP5A1 in isolated heart mitochondria extracts using a ATP5A1 Monoclonal antibody (Product # 43-9800) at a concentration of 1 µg/mL. Lane 1: Isolated mitochondria from human heart at 10 µg, Lane 2: Isolated mitochondria from bovine heart at 4 µg, Lane 3: Isolated mitochondria from rat heart at 10 µg. Lane 4: Isolated mitochondria from mouse heart at 10 µg, Lane 5: HepG2 (Human liver hepatocellular carcinoma cell line) lysate at 20 µg. Predicted band size: 53 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of ATP5A1 was performed using HeLa cells. The cells were fixed with 100% methanol for 10 minutes, permeabilized with 0.1% Triton X-100 for 5 minutes, and blocked with 1% BSA/10% normal goat serum/0.3 M glycine in 0.1% PBS-Tween for 1 hour. The cells were labeled with Tubulin Monoclonal Antibody and ATP5A1 Monoclonal Antibody (15H4C4) (Product # 43-9800) at 1 µg/mL, incubated at 4°C overnight and then incubated for 1 hour with Goat Anti-Mouse IgG (H+L) Secondary Antibody (Alexa Fluor® 647) preadsorbed at 0.5 µg/mL, Goat Anti-Rat IgG (H+L) Secondary Antibody (Alexa Fluor® 488) preadsorbed at 0.5 µg/mL and DAPI.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of ATP5A1 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 ATP5A1 Monoclonal Antibody (15H4C4) (Product # 43-9800) at 2 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) 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). 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: Flow cytometric analysis of ATP5A1 in HepG2 cells using a ATP5A1 Monoclonal Antibody (Product # 43-9800) at 1 µg/1x10^6 cells, shown in red. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) at 1:500 dilution. Isotype control, as seen in black, was a mouse IgG2b antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Partial localization of Dicer and Ago2 in the mitochondrial fraction of WT and MELAS cybrid cells. (A) Immunofluorescence microscopy analysis of endogenous Dicer and Ago2 subcellular localization in WT and MELAS cybrid cells. Nuclei were stained with DAPI (blue), and mitochondria were immunodetected with anti-ATP5A1 (OXPHOS Complex V subunit) or anti-CLPP (a serine protease located in the mitochondrial matrix). Scale bars, 10 mum. (B) Representative immunoblots of Dicer and Ago2 in subcellular fractions of WT and MELAS cybrid cells. Dicer and Ago2 levels were determined in nucleus and intact cells, cytosol and mitoplasts enriched fractions together with respective markers (Pol II (nucleus), Vinculin (cytoplasm), and SDHA (mitochondria)).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 4 Rictor controls HFD-induced mitochondrial localization of Drp1. (A) Immunostaining of mitochondrial localization of Drp1 protein in cardiomyocytes of ND- or HFD-treated control and rictor knockdown flies. Scale bar: 10 mum. (B) The quantification of Pearson correlation coefficient of the colocalization between Drp1 and mitochondrial marker (ATP5A1). Two-way ANOVA: Interaction between diet and genotype is highly significant, p < 0.0001. Tukey's multiple comparison test: *** p < 0.001, ns: not significant. N = 4~5 (4~5 hearts per genotype). (C) Immunostaining of mitochondria and F-actin of cardiomyocytes from ND- or HFD-treated control and Drp1 knockdown flies. Scale bar: 10 mum. (D) The proportion of the elongated mitochondria of the control and Drp1 knockdown flies under ND or HFD treatment. Two-way ANOVA: Interaction between diet and genotype is significant, p = 0.0098. Tukey's multiple comparison test: ** p < 0.01, ns: not significant. N = 4~6 (4~6 hearts per genotype). The control group re-uses the data from Figure 1F . (E) The proportion of the elongated mitochondria of the control, rictor knockdown, and rictor RNAi ; Drp1 OE flies under ND or HFD treatment. Two-way ANOVA: Interaction between diet and genotype is significant, p = 0.0072. Tukey's multiple comparison test: * p < 0.05, ns: not significant. N = 3~4 (3~4 hearts per genotype).

43-9800

Antibody data