Anti-UQCRC2 抗体 [13G12AF12BB11]

• WB

Unknown

Western blot - Anti-UQCRC2 antibody [13G12AF12BB11] (AB14745)

All lanes:

Western blot - Anti-UQCRC2 antibody [13G12AF12BB11] (ab14745) at 0.5 µg/mL

Lane 1:

Human skeletal muscle tissue lysate at 10 µg

Lane 2:

Ramos (human Burkitt's lymphoma cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Goat polyclonal to mouse IgG - H&L - pre-adsorbed (HRP) at 1/3000 dilution

Predicted band size: 48 kDa

Observed band size: 30 kDa,48 kDa

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• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-UQCRC2 antibody [13G12AF12BB11] (AB14745)

IHC image of ab14745 staining in human heart 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 ab14745, 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.

• Flow Cyt

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Flow Cytometry - Anti-UQCRC2 antibody [13G12AF12BB11] (AB14745)

Overlay histogram showing HepG2 cells stained with ab14745 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab14745, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1](ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• IF

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Immunofluorescence - Anti-UQCRC2 antibody [13G12AF12BB11] (AB14745)

Mitochondrial localization of complex III visualized by immunofluorescence using anti-complex III subunit Core 2 mAb 2E3GC12FB2AE2 (ab14745). Cultured human embryonic lung-derived fibroblasts (strain MRC5) were fixed, permeabilized and then labeled with ab14745 (0.5 μg/ml) followed by an AlexaFluor® 488-conjugated-goat-anti-mouse IgG1 isotype specific secondary antibody (2 μg/ml).

• WB

AbReview41073****

Western blot - Anti-UQCRC2 antibody [13G12AF12BB11] (AB14745)

All lanes:

Western blot - Anti-UQCRC2 antibody [13G12AF12BB11] (ab14745) at 1/1000 dilution

All lanes:

H23 whole cell lysate at 20 µg

Secondary

All lanes:

HRP-conjugated goat anti-mouse IgG polyclonal at 1/2000 dilution

Predicted band size: 48 kDa

Observed band size: 48 kDa

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Exposure time: 1min

This image is courtesy of an anonymous Abreview

• WB

CiteAb

Western blot - Anti-UQCRC2 antibody [13G12AF12BB11] (AB14745)

Western Blotting using Anti-UQCRC2 antibody [13G12AF12BB11], ab14745. Publication image from Protasoni, M. et al., 2020, EMBO J, 31912925. Legend direct from paper.

Blue‐native gel electrophoresis (BNGE) mass spectrometry and immunodetection analysis of cIII2‐related proteinsComplexome profiles of cIII2 structural subunits generated by analyzing the peptide content in each of the 64 slices in which the gel lanes were excised (see also Figs EV1 and EV2). The graphs plot the relative peptide peak intensities along the lane, setting the maximum to 1.0, versus the molecular mass calculated using the individual complexes and supercomplexes as the standards to generate a calibration curve. The relative amounts of the proteins between the two cell lines were determined by calculating the H/L ratios of peptides that were present in both WT (blue traces) and δ4‐CYB samples (red traces). The represented values are the mean ± SEM of the two reciprocal labeling experiments.Second‐dimension BNGE of digitonin‐solubilized samples from WT and δ4‐CYB cells, Western blot and immunodetection of the indicated cIII2 structural subunits with specific antibodies. The immunodetection patterns were equivalent to the complexome profiles.Quantification of the total peak area under the curves (AUC) defined by the peptide intensity peaks for the indicated cIII2 subunits. The x‐axis values were the slice number (1‐64), and the y‐axis values were the relative peptide intensity. The graph shows the mean ± SD (n = 2). Two‐way ANOVA with Sidak's multiple comparisons test **P = 0.0083 (UQCRC2); **P = 0.0033 (UQCRFS1); *P = 0.0224; n.s. = non‐significant.Complexome profiles of two cIII2 assembly factors (BCS1L and LYRM7 or MZM1L) generated in the same way as in (A). The represented values are the mean ± SEM of the two reciprocal labeling experiments.Source data are available online for this figure.

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• WB

CiteAb

Western blot - Anti-UQCRC2 antibody [13G12AF12BB11] (AB14745)

Western Blotting using Anti-UQCRC2 antibody [13G12AF12BB11], ab14745. Publication image from Kopajtich, R. et al., 2017, Nat Commun, 28604674. Legend direct from paper.

RNA aberrant expression detection and validation.(a) Aberrantly expressed genes (Hochberg corrected P value<0.05 and |Z-score|>3) for each patient fibroblasts. (b) Gene-wise RNA expression volcano plot of nominal P values (−log10P value) against Z-scores of the patient #35791 compared against all other fibroblasts. Z-scores with absolute value >5 are plotted at ±5, respectively. (c) Same as (b) for patient #73804. (d) Sample-wise RNA expression is ranked for the genes TIMMDC1 (top) and MGST1 (bottom). Samples with aberrant expression for the corresponding gene are highlighted in red (#35791, #66744, and #73804). (e) Gene-wise comparison of RNA and protein fold changes of patient #35791 compared to the average across the fibroblast cell lines of all other patients. Subunits of the mitochondrial respiratory chain complex I are highlighted (red squares). Reliably detected proteins that were not detected in this sample are shown separately with their corresponding RNA fold changes (points below solid horizontal line). (f) Western blot of TIMMDC1, NDUFA13, NDUFB3 and NDUFB8 protein in three fibroblast cell lines without (#62346, #91324, NHDF) and three with a variant in TIMMDC1 (#35791, #66744 and #96687), and fibroblasts re-expressing TIMMDC1 (‘-T’) (#35791-T, #66744-T and #96687-T). UQCRC2 was used as loading control. CI, complex I subunit; CIII, complex III subunit; MW, molecular weight. (g) Blue native PAGE blot of the control fibroblasts re-expressing TIMMDC1 (NHDF-T), the control fibroblasts (NHDF), patient fibroblasts (#96687) and patient fibroblast re-expressing TIMMDC1 (#96687-T). Immunodecoration for complex I and complex III was performed using NDUFB8 and UQCRC2 antibodies, respectively. CI, complex I subunit; CIII, complex III subunit.

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