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Rabbit anti keratin 10

Manufactured by BioLegend
Sourced in Canada, United Kingdom, United States

Rabbit anti-Keratin 10 is an antibody that recognizes the keratin 10 protein. Keratin 10 is a structural protein found in the intermediate filaments of epithelial cells. This antibody can be used for the detection and localization of keratin 10 in various applications.

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3 protocols using rabbit anti keratin 10

1

Western Blot Analysis of Cell Signaling

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Whole cell lysates for Western Blot were prepared with the SDS-sample buffer. Proteins were separated by SDS-PAGE and transferred onto the PVDF membrane (Millipore/Fisher Scientific, Pittsburg, PA), Darmstadt, Germany). Antibodies for immunoblotting from Cell Signaling Technology (Dancers, MA) would include rabbit anti-Bmal1 (#14020), rabbit anti-CLOCK (#5157), rabbit anti-phospho-p53 (Ser15) [#9284], rabbit anti-p21 Waf1/Cip1 (#2947), rabbit anti-Histone H2AX (#2595), rabbit anti-cleaved PARP (#9541), and rabbit anti-phospho-CHK1 (Ser 345) (#2348). Other antibodies include mouse anti-p53 (sc-126; Santa Cruz Biotechnology Inc., Santa Cruz, CA), rabbit anti-Keratin 10 (Poly19054; Biolegend, San Diego, CA), mouse anti-phospho-Histone H2AX (Ser139) (05–636; Millipore/Fisher Scientific), and mouse anti-α-tubulin (T9026; Sigma-Aldrich, St. Louis, MO). HRP-conjugated secondary antibodies were from Santa Cruz Biotechnology Inc. Chemiluminescence images were acquired using an Amersham Imager 600 from GE Healthcare Life Sciences (Pittsburgh, PA) or iBright FL1000 (Invitrogen-Life Technology/Fisher Scientific, Pittsburgh, PA). The level of target proteins was quantified by densitometry scanning with the ImageJ software and normalized to the amount of α-tubulin.
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2

Immunofluorescence Analysis of Tight Junction Proteins

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Skin and buccal mucosa specimens were frozen in an optimal cutting temperature (OCT) compound (Cryomount I, 33351; Muto Pure Chemicals, Japan) and sectioned with a cryostat (Leica CM1950, Leica Biosystems, Germany). To prepare horizontal‐sliced sections, specimens were embedded horizontally in the OCT compound, and 5 μm‐thick sections were prepared. Frozen sections were incubated with primary antibodies: rabbit anti‐ZO‐1 (dilution 1:200 in PBS, ab2168; Abcam, UK), rabbit anti‐claudin‐1 (dilution 1:200 in PBS, ab15098; Abcam, UK), rabbit anti‐occludin (dilution 1:100 in PBS, 71‐1500; Invitrogen, CA), mouse anti‐keratin14 (dilution 1:200 in PBS, ab7800; Abcam, UK), rabbit anti‐keratin10 (dilution 1:500 in PBS, 905404; BioLegend, CA), rabbit anti‐cleaved caspase‐3 (dilution 1:100 in PBS, #9661 S, Cell Signalling Technology, MA) or anti‐Ki‐67 (dilution 1: 200, NB600‐152, Novus Biologicals) at 37°C for 60 min. The sections were subsequently incubated with Fluorescein isothiocyanate (FITC)‐conjugated goat anti‐rabbit IgG H + L (dilution 1:200 in PBS, 234; MBL), alexa 488‐conjugated anti‐mouse IgG3 (dilution 1:1000 in PBS) or alexa 568‐conjugated anti‐rabbit IgG (dillution 1:1000 in PBS) at 37°C for 60 min and mounted in a Pharma Fluor aqueous mounting medium (TA‐006‐FM; Thermo Fisher Scientific, MA). The stained immunofluorescent samples were observed with a BZ9000 microscope.
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3

Antibody Validation for Cellular Markers

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The following primary antibodies were used in this study: mouse anti-Flag (#F1804, 1:5000, Sigma-Aldrich), mouse and goat anti-HA (#901502, Biolegend, San Diego, CA, USA; #NB600–362, Novus, St. Louis, MO, USA), anti-V5 tag antibody (Invitrogen), chicken anti-Keratin5 (#905904, 1:500, Biolegend), rabbit anti-Keratin10 (#905404, 1:500, Biolegend), rabbit anti-Keratin6A (#19057, 1:500, Biolegend).
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