Rabbit anti k14

Manufactured by Fortrea

Rabbit anti-K14 is a polyclonal antibody that specifically binds to the K14 (Keratin 14) protein. K14 is a type I intermediate filament protein found in the basal layer of stratified squamous epithelial tissues. This antibody can be used for the detection and analysis of K14 expression in various research applications.

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Anti-K14

4 protocols using rabbit anti k14

Immunohistochemical Analysis of Skin

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Samples were processed as previously described (Ilic et al., 2007; Stephenson et al., 2012 ). The following antibodies were used: rabbit anti-K14, rabbit anti-filaggrin, rabbit anti-involucrin, and rabbit anti-loricrin (all from Covance); rabbit anti-laminin (DAKO); mouse anti-p63 (Santa Cruz Biotechnology); rabbit anti-fibronectin, mouse anti-collagen IV, mouse anti-collagen VII, and rabbit anti-desmocollin 1 (all from Sigma-Aldrich); and rabbit anti-p63, mouse anti-K10, and rabbit anti-collagen I (all from Abcam). Secondary antibodies were obtained from Jackson ImmunoResearch and Life Technologies.

Petrova A., Celli A., Jacquet L., Dafou D., Crumrine D., Hupe M., Arno M., Hobbs C., Cvoro A., Karagiannis P., Devito L., Sun R., Adame L.C., Vaughan R., McGrath J.A., Mauro T.M, & Ilic D. (2014). 3D In Vitro Model of a Functional Epidermal Permeability Barrier from Human Embryonic Stem Cells and Induced Pluripotent Stem Cells. Stem Cell Reports, 2(5), 675-689.

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Immunostaining of Skin and Cell Cultures

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Skin cryosections or cultured cells were fixed in 4% paraformaldehyde for 1 h and permeabilized in 0.25% Triton X‐100 for 15 min at room temperature, blocked in 1% BSA for 30 min at room temperature and then stained with Mouse anti‐MyHC (Millipore, 05–716), Rabbit anti‐K14 (Covance, 905301), Mouse anti‐Sox10 (Santa Cruz Biotechnology, SC‐365692), Mouse anti‐Runx3 (R&D Systems, MAB3765‐SP), Rabbit anti‐Mitf (Thermo Fisher, MA5‐32554), Mouse anti‐Pax7 (DSHB), and Rabbit anti‐Laminin (Sigma, L9393) at 4°C overnight with gentle rocking, followed by Alexa 488‐, 561‐ or 647‐labelled anti‐mouse or anti‐rabbit secondary antibody (Invitrogen) staining at room temperature for 1 h. Nuclei were stained with 5 mg/mL DAPI (Invitrogen) for 5 min. All images were acquired by confocal microscopy (Leica). For Pax7 staining, heat‐activated antigen retrieval was performed by placing the paraformaldehyde‐fixed samples in citrate buffer (pH 6.0) at 95°C for 20 min and cooling the slides at room temperature for 20 min, followed by antibody incubation as described above.

Yang W., Wang Y., Du Y., Li J., Jia M., Li S., Ma R., Li C., Deng H, & Hu P. (2023). Chemical reprogramming of melanocytes to skeletal muscle cells. Journal of Cachexia, Sarcopenia and Muscle, 14(2), 903-914.

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Immunodetection of Wnt4 and Keratins

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Rabbit polyclonal anti-Wnt4 antibody was raised against a synthetic peptide (NH2-PRNAQFKPHTDEDC-CONH2) and affinity-purified. Other antibodies used were rat anti K8 (1:250; Developmental Hybridoma Bank), rabbit anti-K14 (1:1000; Covance), rabbit anti-Flag (1:1000; Sigma), mouse anti-β-Actin (1:3000; Sigma), and rabbit anti-milk (1:500; Nordic Immunological Laboratories).

Cai C., Yu Q.C., Jiang W., Liu W., Song W., Yu H., Zhang L., Yang Y, & Zeng Y.A. (2014). R-spondin1 is a novel hormone mediator for mammary stem cell self-renewal. Genes & Development, 28(20), 2205-2218.

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Immunofluorescence and In Situ Hybridization

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Frozen sections were prepared by air-drying and fixation for 1 h in cold MeOH. Tissue sections were incubated with primary antibodies overnight at 4°C followed by washes, incubation with secondary antibodies for 2 h at 25°C, and counterstaining with DAPI (Vector Laboratories). The following antibodies were used: rat anti-K8 (1:250; Developmental Hybridoma Bank) and rabbit anti-K14 (1:1000; Covance). In situ hybridization was performed using the RNAscope kit (Advanced Cell Diagnostics) following the manufacturer’s instructions. Wnt4, Wnt7B, Rspo1, and PR probes were ordered from Advanced Cell Diagnostics. For all of the immunofluorescence stainings, in situ and in immunoblots, at least three independent experiments were conducted. Representative images are shown in the figures.

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