Anti-Cytochrome C antibody

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Cytochrome C antibody (AB90529)

ab90529 staining cytochrome C in HepG2 cells treated with mevastatin sodium salt (ab120854), by ICC/IF. Increase of cytochrome C expression correlates with increased concentration of mevastatin sodium salt, as described in literature.
The cells were incubated at 37°C for 4 hours in media containing different concentrations of ab120854 (mevastatin sodium salt) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab90529 (1 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Cytochrome C antibody (AB90529)

ab90529 staining cytochrome C in HepG2 cells treated with mevastatin (ab120652), by ICC/IF. Increase of cytochrome C expression correlates with increased concentration of mevastatin, as described in literature.
The cells were incubated at 37°C for 4 hours in media containing different concentrations of ab120652 (mevastatin) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab90529 (1 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome C antibody (AB90529)

IHC image of Cytochrome C staining in human skeletal muscle formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab90529, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Cytochrome C antibody (AB90529)

ab90529 staining Cytochrome C in HepG2 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab90529 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• WB

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Western blot - Anti-Cytochrome C antibody (AB90529)

All lanes:

Western blot - Anti-Cytochrome C antibody (ab90529) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

Lane 4:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

Lane 5:

Human testis tissue lysate - total protein (<a href='/en-us/products/unavailable/human-testis-tissue-lysate-total-protein-ab30257'>ab30257</a>) at 10 µg

Lane 6:

Human liver tissue lysate - total protein (<a href='/en-us/products/unavailable/human-liver-tissue-lysate-total-protein-ab29889'>ab29889</a>) at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 11 kDa

Observed band size: 15 kDa

true

Exposure time: 30min

• WB

Unknown

Western blot - Anti-Cytochrome C antibody (AB90529)

All lanes:

Western blot - Anti-Cytochrome C antibody (ab90529) at 1 µg/mL

Lane 1:

Heart (Mouse) Tissue Lysate at 10 µg

Lane 2:

Heart (Rat) Tissue Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 11 kDa

Observed band size: 14 kDa,30 kDa,65 kDa

true

Exposure time: 4min

• WB

CiteAb

Western blot - Anti-Cytochrome C antibody (AB90529)

Cytochrome C western blot using anti-Cytochrome C antibody ab90529. Publication image and figure legend from Ding, X., Sun, W., et al., 2018, Cancer Cell Int, PubMed 30459526.

ab90529 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab90529 please see the product overview.

IL-2 activated the mitochondrial apoptotic pathway in the presence of sorafenib. a, b Mitochondrial ROS production was detected in HepG2 cells. IL-2 (5 ng/ml) treatment was carried out in the presence of 5 μM sorafenib. c–e The antioxidants in HepG2 cells under IL-2 and sorafenib co-treatment were measured via ELISA. f The mPTP opening rate was analysed to determine the mitochondrial damage. IL-2 (5 ng/ml) treatment was carried out in the presence of 5 μM sorafenib. g, h Cyt-c liberation was observed via immunofluorescence. i–o Mitochondrial apoptotic proteins were analysed by western blot. The sorafenib-mediated upregulation of apoptotic proteins was further augmented by IL-2 treatment. *p < 0.05 vs. control group; #p < 0.05 vs. sorafenib group. Cont control

false