Anti-Cytochrome b5 antibody

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytochrome b5 antibody (AB69801)

IHC image of ab69801 staining in human normal liver 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 ab69801, 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

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Immunocytochemistry/ Immunofluorescence - Anti-Cytochrome b5 antibody (AB69801)

Left : NIH/3T3 cells stained with ab69801 at 1/500 dilution; Right : the antibody was pre-incubated with the immunizing peptide.

• WB

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Western blot - Anti-Cytochrome b5 antibody (AB69801)

All lanes:

Western blot - Anti-Cytochrome b5 antibody (ab69801) at 1/500 dilution

Lane 1:

extracts from A549 cells at 15 µg

Lane 2:

extracts from A549 cells at 15 µg with immunizing peptide

Predicted band size: 15 kDa

Observed band size: 15 kDa

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• WB

CiteAb

Western blot - Anti-Cytochrome b5 antibody (AB69801)

Cytochrome b5 western blot using anti-Cytochrome b5 antibody ab69801. Publication image and figure legend from Khan, R., Zahid, S., et al., 2013, Clin Proteomics, PubMed 23724895.

ab69801 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 ab69801 please see the product overview.

Immunoblot analysis confirms changes in protein expression of CYB5A. (A) : representative western blot analysis of CYB5A in HCC, fibrotic liver and HepG2 cell line. For this, 100μg of nuclear membrane samples were loaded onto a 12% SDS-PAGE. Expression level was analyzed using the primary antibody CYB5A (1 : 1000 dilution) and horse raddish peroxidase (HRP) - conjugated secondary antibody (1 : 5000 dilution). β-actin and GAPDH were used as a loading control. (B) : representative graphical expression of validated proteins by western blot. Digital images were taken by gel documentation system (Bio-Rad). Quantification and intensity measurement of protein bands were analyzed by Quantity One gel analysis software (Bio-Rad). Statistical significance (p- value > 0.05) was calculated using SPSS statistics version 17.

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