Anti-CLPX antibody [EP8772]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CLPX antibody [EP8772] (AB168338)

Immunofluorescence analysis of A673 cells (red) labeling CLPX with ab168338 at 1/100 dilution. DAPI nuclear staining (blue).

• WB

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Western blot - Anti-CLPX antibody [EP8772] (AB168338)

All lanes:

Western blot - Anti-CLPX antibody [EP8772] (ab168338) at 1/1000 dilution

Lane 1:

A673 lysates at 10 µg

Lane 2:

HT-1080 lysates at 10 µg

Lane 3:

Saos-2 lysates at 10 µg

Lane 4:

Human fetal liver lysates at 10 µg

Lane 5:

Human fetal heart lysates at 10 µg

Predicted band size: 69 kDa

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• WB

CiteAb

Western blot - Anti-CLPX antibody [EP8772] (AB168338)

CLPX western blot using anti-CLPX antibody [EP8772] ab168338. Publication image and figure legend from Franco-Iborra, S., Cuadros, T., et al., 2018, Cell Death Dis, PubMed 30405116.

ab168338 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 ab168338 please see the product overview.

Complex I inhibition leads to a mitochondrial dysfunction and cell death.a–f BE(2)-M17 cells were treated with MPP+ (1 mM, 24 h) or untreated (UT). a Representative immunoblots of ATP5A (complex V), UQCRC2 (complex III), SDHB (complex II), COX II (complex IV), NDUFB8 (complex I) and VDAC protein levels normalized relative to VDAC (n = 3 independent experiments). Quantification is depicted as fold change to UT condition (*P < 0.05, **P < 0.01, ***P < 0.001 compared with UT condition after unpaired two-sided Student's t-test). b Representative immunoblots of SDHB and ATP5A protein levels in soluble and insoluble mitochondria-isolated fractions (n = 3 independent experiments). Quantification is depicted as fold change to UT condition (*P < 0.05, **P < 0.01 after unpaired two-sided Student's t-test). cCLPX and HSP9 gene expression levels normalized relative to RPLP0 (n = 3 independent experiments). Quantification is depicted as fold change to UT condition (**P < 0.01, ***P < 0.001 after unpaired two-sided Student's t-test). d Representative immunoblots of CLPX, GRP75 and VDAC protein levels normalized relative to VDAC (n = 4 independent experiments). Quantification is depicted as fold change to UT condition (*P < 0.05 after unpaired two-sided Student's t-test). e Mitochondrial membrane potential measured as TMRM fluorescence intensity. Quantification is depicted as the fold change in fluorescence intensity compared with UT condition (n = 3 independent experiments, ***P < 0.001 after unpaired two-sided Student's t-test). f ROS production measured as CM-H2DCFDA fluorescence intensity. Quantification is depicted as the fold change in fluorescence intensity compared with UT condition (n = 3 independent experiments, **P < 0.01 after unpaired two-sided Student's t-test). g In vitro sensitivity of BE(2)-M17 cells to increasing MPP+ concentrations for 24 h (n = 4 independent experiments). Cell survival was determined by MTT assay. Quantification is depicted as the % of cell survival relative to UT condition (*P < 0.05 after one-way ANOVA test followed by Tukey's post hoc test). Error bars indicate s.e.m.

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