Rabbit anti keratin 14

Manufactured by Fortrea

Rabbit anti-Keratin 14 is a primary antibody that specifically binds to the keratin 14 protein. Keratin 14 is a member of the keratin family of intermediate filament proteins and is commonly used as a marker for basal cells in epithelial tissues.

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Lab products found in correlation

Rabbit anti keratin 10, by Fortrea (2 mentions) Alexa fluor 488 goat anti rabbit, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Prolong gold mounting medium, by Thermo Fisher Scientific (1 mentions) Rabbit anti involucrin, by Abcam (1 mentions) Tcs sp5 confocal laser scanning microscope, by Leica (1 mentions) Dmi8 inverted microscope, by Leica (1 mentions) Alexa fluor 488 goat anti mouse, by Thermo Fisher Scientific (1 mentions) Alexa fluor 555 goat anti rabbit, by Thermo Fisher Scientific (1 mentions) Mouse anti p63, by Santa Cruz Biotechnology (1 mentions) Eclipse 90i fluorescent microscope, by Nikon (1 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions) Rabbit anti loricrin, by Fortrea (1 mentions)

Close spelling variants of this product

Rabbit anti-human keratin 14 (AF64; Anti–keratin 14 Keratin 14 Rabbit anti-keratin

3 protocols using rabbit anti keratin 14

Immunostaining of Skin Sections

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For immunostaining of skin sections in Figure 1—figure supplement 1, dorsal skin was prepared and sectioned as for X-Gal staining. Skin cryosections were fixed for 10 min in cold acetone, washed twice in 1× PBS prior to blocking for 30 min in 10% normal goat serum at room temperature. Primary antibodies used were Rabbit anti-Keratin 14 (Covance, Cat#PRB-155P), 1:1000 (1 µg/ml), and Rabbit anti-Keratin 10 (Covance, Cat#PRB-159P), 1:1000 (1 µg/ml). Secondary antibody used was Goat anti-Rabbit Alexa Fluor 488 (Invitrogen, Cat#A11008), 1:1000. Nuclei were stained by DAPI (Invitrogen, Cat#D1306), 1:50,000. All antibody incubations were performed at room temperature, for 1 hr in 1% BSA in PBS. Slides were mounted in gelvatol containing DAPI.

Holt J.R., Zeng W.Z., Evans E.L., Woo S.H., Ma S., Abuwarda H., Loud M., Patapoutian A, & Pathak M.M. (2021). Spatiotemporal dynamics of PIEZO1 localization controls keratinocyte migration during wound healing. eLife, 10, e65415.

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Immunofluorescence Staining of Stem Cells

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Cells were fixed (4% paraformaldehyde), permeabilized (0.2% Triton X-100; except for Fig. 2H – TRA-1-81), blocked (BSA), and stained with antibodies against as per standard laboratory procedures. Nuclei were counterstained with DAPI. Images were acquired with a Leica DMi8 inverted microscope. Primary antibodies used: TRA-1-81 (Sigma Aldrich MAB4381), TRA-1-60 (Sigma Aldrich MAB4360), NANOG (Abcam ab21624), K18 (1:800, R&D AF7619), K14 (1:800, BioLegend SIG-3476-100), and p63 (1:100 Gene Tex GTX102425), ITGA6 (1:200, Millipore, MAB1378). Antibodies for mouse grafts comprised of rabbit anti-Involucrin (1:100, abcam), rabbit anti-keratin 14 (1:2000, Covance), mouse anti-keratin 18 (1:800 Abcam), rabbit anti-keratin 10 (1:500, Covance), human specific N terminal anti-collagen VII LH7.2 (1:250 Millipore), The fluorescence images were taken using the TCS SP5 confocal laser scanning microscope (Leica). Tissue sections were co-stained with 1:10000 Hoechst for 10 min. and slides were mounted with the Prolong Gold mounting medium (Life Technologies).

Neumayer G., Torkelson J.L., Li S., McCarthy K., Zhen H.H., Vangipuram M., Jackow J., Rami A., Hansen C., Guo Z., Gaddam S., Pappalardo A., Li L., Cramer A., Roy K.R., Nguyen T.M., Tanabe K., McGrath P.S., Bruckner A., Bilousova G., Roop D., Bailey I., Tang J.Y., Christiano A., Steinmetz L.M., Wernig M, & Oro A.E. (2023). A scalable, GMP-compatible, autologous organotypic cell therapy for Dystrophic Epidermolysis Bullosa. bioRxiv.

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Immunofluorescent Detection of Epidermal Markers

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For immunofluorescent detection of markers of the epidermis, sections were deparaffinized and rehydrated in reducing concentrations of ethanol. Sections were subjected to antigen retrieval by boiling in 10mM Sodium citrate (pH6.0) for 30 minutes. Tissues sections were permeabilized using 0.5% Triton X-100 followed by blocking in blocking solution (10% normal goat serum, 0.1% Bovine Serum Albumin in 1X Phosphate Buffered Saline (PBS)) for one hour at room temperature. Sections were then incubated in anti-mouse F′ (ab) fragment (Jackson ImmunoResearch Laboratories, West Grove, PA) for five minutes to reduce non-specific binding of anti-mouse secondary. Sections were incubated with 1:100 rabbit anti-Irf6 (Sigma-Aldrich; St. Louis, MO), 1:250 rabbit anti-Keratin 14 (Covance), 1:150 mouse anti-p63 (Santa Cruz), and 1:250 rabbit anti-loricrin (Covance) overnight at 4°C, followed by incubation with either goat anti-mouse AlexaFluor 488 or goat anti-rabbit AlexaFluor 555 (Life Technologies, Grand Island, NY). Nuclei were counterstained for 10 minutes in a 1:10,000 dilution of 4′ 6-diaminidino-2-phenylindole (DAPI; Life Technologies, Grand Island, NY). Slides were then mounted in ProLong GOLD Antifade reagent (Life Technologies, Grand Island, NY). Images were taken using a Nikon Eclipse 90i fluorescent microscope.

Smith A.L., Kousa Y.A., Kinoshita A., Fodor K., Yang B, & Schutte B.C. (2017). Generation and Characterization of a Conditional Allele of Interferon regulatory factor 6. Genesis (New York, N.Y. : 2000), 55(7), 10.1002/dvg.23038.

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