Anti-AKR1C1 antibody

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-AKR1C1 antibody (AB192785)

Immunofluorescence analysis of A431 cells, labeling AKR1C1 (green, upper panel) with ab192785 at 1/200 dilution. Nuclei were counterstained with Hoechst 33342 (blue, lower panel).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AKR1C1 antibody (AB192785)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded human liver tissue, labeling AKR1C1 with ab192785 at 7.5 μg/ml.

• WB

Supplier Data

Western blot - Anti-AKR1C1 antibody (AB192785)

10% SDS PAGE

All lanes:

Western blot - Anti-AKR1C1 antibody (ab192785) at 1/1000 dilution

All lanes:

Raji whole cell lysate at 30 µg

Predicted band size: 37 kDa

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• WB

Supplier Data

Western blot - Anti-AKR1C1 antibody (AB192785)

10% SDS-PAGE

All lanes:

Western blot - Anti-AKR1C1 antibody (ab192785) at 1/1000 dilution

All lanes:

mouse liver tissue lysate at 50 µg

Predicted band size: 37 kDa

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• WB

CiteAb

Western blot - Anti-AKR1C1 antibody (AB192785)

AKR1C1 western blot using anti-AKR1C1 antibody ab192785. Publication image and figure legend from Huppke, P., Weissbach, S., et al., 2017, Nat Commun, PubMed 29018201.

ab192785 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 ab192785 please see the product overview.

Increased stabilization and activation of mutant NRF2. a Representative western blot of endogenous level of NRF2, KEAP1, G6PD, AKR1B10 and AKR1C1 in protein lysates of human primary fibroblast cell lines from two controls (NRF2 WT 1, WT 2) and patient 1 with NRF2 p.T80K variant. Full blots are shown in Supplementary Fig. 6. b Quantitative analysis of western blot images illustrating the endogenous level of NRF2, KEAP1, G6PD, AKR1B10 and AKR1C1 relative normalized to ACTB and NRF2 WT 1. c qRT–PCR analysis of NFE2L2, KEAP1 and target gene expression in primary fibroblast cell lines from two controls (NRF2 WT 1, WT 2) and patient 1 with NRF2 p.T80K variant. AKR1B10 and AKR1C1 are visualized on a separated X axis due to the high range. Expression is normalized to that of ACTB. % of mRNA is equal to 2−∆∆CT and normalized relative to NRF2 WT 1. Redox calibration confirms full functionality of roGFP1 as well as identical response ranges for NRF2 WT 2 and NRF2 p.T80K fibroblast cells. d Response range calibration of an exemplary NRF2 WT 2 and NRF2 p.T80K fibroblast cell performed as a continuous recording of the roGFP1 ratio F395/F470 within a ROI of cytoplasm of the cell, scale bar is 20 µM. Plotted traces represent full oxidation (Rox, induced by 5 mM H2O2, 5 min) and full reduction (Rred, induced by 10 mM DTT, 5 min). After calibration the relative degrees of roGFP1 oxidation and corresponding roGFP1 redox potentials can be calculated. e Baseline redox conditions of NRF2 WT 2 and NRF2 p.T80K fibroblasts. Upper diagram shows the relative level of roGFP1 oxidation of NRF2 WT 2 and NRF2 p.T80K cells at rest (OxDroGFP1, Eq. 1). Lower diagram represents corresponding steady-state roGFP1 redox potential (EroGFP1, Eq. 2). b, c Data are given as means ± SEM, n ≥ 3 independent experiments. Data were analyzed by one-way analysis of variance with multiple comparisons : *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. e Data are given as means ± SEM. Number of measured cells are given within the bar. Statistical differences were obtained with unpaired Welch’s t-test : ***p ≤ 0.001

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