Anti-HSD17B1 antibody [EP1682Y]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-HSD17B1 antibody [EP1682Y] (AB51045)

Overlay histogram showing JEG3 cells stained with ab51045 (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. The cells were then incubated with the antibody (ab51045, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HSD17B1 antibody [EP1682Y] (AB51045)

ab51045 (1/100 dilution) staining human HSD17B1 in paraffin embedded human placenta tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-HSD17B1 antibody [EP1682Y] (AB51045)

Purified ab51045 at 1/50 dilution (2μg) immunoprecipitating HSD17B1 in Human placenta lysate.
Lane 1 (input) : Human placenta lysate 10μg
Lane 2 (+) : ab51045 + Human placenta lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab51045 in Human placenta lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 34 kDa

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Immunoprecipitation - Anti-HSD17B1 antibody [EP1682Y] (ab51045)

Predicted band size: 35 kDa

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• WB

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Western blot - Anti-HSD17B1 antibody [EP1682Y] (AB51045)

All lanes:

Western blot - Anti-HSD17B1 antibody [EP1682Y] (ab51045) at 1/100000 dilution

All lanes:

Human placenta tissue lysate at 10 µg

Secondary

All lanes:

Goat anti-Rabbit HRP labeled at 1/2000 dilution

Predicted band size: 35 kDa

Observed band size: 35 kDa

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• WB

CiteAb

Western blot - Anti-HSD17B1 antibody [EP1682Y] (AB51045)

HSD17B1 western blot using anti-HSD17B1 antibody [EP1682Y] ab51045. Publication image and figure legend from Aka, J. A., Zerradi, M., et al., 2012, Breast Cancer Res, PubMed 22691413.

ab51045 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 ab51045 please see the product overview.

Proteomic analysis of wild type (WT) MCF7 cells and MCF7 cells stably transfected with 17β-HSD1 (MCF7-17βHSD1). (A) 17beta-hydroxysteroid dehydrogenase type 1 (17β-HSD1) and β-actin expression between WT MCF7 and MCF7-17βHSD1 revealed by western blots. (B) Representative two-dimensional gel images for WT MCF7 and MCF7-17βHSD1 cells. Whole cell lysates (200 μg) from each cell were separated by two-dimensional electrophoresis and visualized by Sypro Ruby staining. The two-dimensional gels were scanned and the differentially expressed (2-fold or higher, p < 0.05) proteins were detected using Progenesis software. The 18 differentially expressed protein spots that were selected for mass spectrometry (MS) analysis are marked with circles. Protein spots upregulated in MCF7-17βHSD1 are depicted in the MCF7-17βHSD1 proteome image; protein spots downregulated in MCF7-17βHSD1 are depicted in the MCF7 proteome image. The numbers refer to the spot number listed in Table 1 and Additional file 2. The squares represent the indicated area shown in more detail in (D). (C) Summary of the numbers of spots and proteins obtained from the proteomics data. *Upregulated (up) and downregulated (down) proteins in MCF7-17βHSD1 as compared to WT MCF7 cells. (D) Zoom showing some differentially expressed protein spots from WT MCF7 and MCF7-17βHSD1 comparison. Arrows indicate 17β-HSD1 protein which was revealed by MS analysis to be present in the spot numbers 2,305 (unique to MCF7-17βHSD1) and 2,300 (upregulated in MCF7-17βHSD1 as compared to WT MCF7 cells).

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