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Anti keratin 14

Manufactured by Thermo Fisher Scientific

Anti-Keratin 14 is a laboratory reagent used for the detection and identification of keratin 14 in cells and tissues through immunochemical techniques. It is a monoclonal antibody that specifically binds to keratin 14, a structural protein found in epithelial cells.

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3 protocols using anti keratin 14

1

Immunostaining of Mammary Gland Tissue

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Mammary glands were fixed in formalin and embedded in paraffin for immunostaining. Sections were de-paraffinized, dehydratated, and microwaved for 10 min at 95 °C in Tris–EDTA (pH 9) for antigen retrieval. The tissue sections were incubated o/n at 4 °C with primary antibodies diluted in PBS + 5% BSA. The samples were incubated with anti-GFP Alexa-488 (1:500), anti-Keratin-14 (1:100), anti-Keratin-8 (1:100), anti-rabbit and anti-rat Alexa-594 conjugated secondary antibodies (Invitrogen) at 1:500 in PBS + 5% BSA 1 h at RT. All the immunofluorescence sections and cells were mounted in ProLong Gold with DAPI. Images were acquired by Carl Zeiss LSM 510 Meta confocal microscope. Images were processed using ImageJ.
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2

Multicolor Immunohistochemistry of Murine Skin

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Skin tissues were embedded in paraffin or snap frozen in OCT compound. Antigen retrieval for paraffin sections was performed in citrate buffer, pH6 for the skin sections. Anti-F4/80 (clone A3-1, MCA497G, BIO-RAD), anti-Keratin 14 (MA5-11599, Invitrogen), anti-Keratin 6 (905701, Biolegend), anti-Keratin 10 (905401, Biolegend) were used for the staining. Alexa-488, Alexa-594 and Alexa-633 fluorescence conjugated secondary antibodies were used for detection. F4/80 staining was performed on cryo sections. All sections were counterstained with DAPI. Quantification of epidermal thickness was performed by measurement of epidermal thickness in five optical fields per section. In each field, four measurements were performed. Percentage of inflamed area was determined as the percentage of inflamed versus total number of optical fields at 20x on individual skin sections. Assessment of tissue pathology was performed in a blinded fashion. All images were acquired using either a Zeiss Meta 710 confocal or PerkinElmer Spinning Disc confocal microscope for fluorescent images and brightfield images were acquired using a Leica SCN400 slide scanner or a Leica DM5500 B microscope.
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3

Multicolor Immunohistochemistry of Murine Skin

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Skin tissues were embedded in paraffin or snap frozen in OCT compound. Antigen retrieval for paraffin sections was performed in citrate buffer, pH6 for the skin sections. Anti-F4/80 (clone A3-1, MCA497G, BIO-RAD), anti-Keratin 14 (MA5-11599, Invitrogen), anti-Keratin 6 (905701, Biolegend), anti-Keratin 10 (905401, Biolegend) were used for the staining. Alexa-488, Alexa-594 and Alexa-633 fluorescence conjugated secondary antibodies were used for detection. F4/80 staining was performed on cryo sections. All sections were counterstained with DAPI. Quantification of epidermal thickness was performed by measurement of epidermal thickness in five optical fields per section. In each field, four measurements were performed. Percentage of inflamed area was determined as the percentage of inflamed versus total number of optical fields at 20x on individual skin sections. Assessment of tissue pathology was performed in a blinded fashion. All images were acquired using either a Zeiss Meta 710 confocal or PerkinElmer Spinning Disc confocal microscope for fluorescent images and brightfield images were acquired using a Leica SCN400 slide scanner or a Leica DM5500 B microscope.
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