Keratin 10

Manufactured by Fortrea

Keratin-10 is a laboratory-grade protein used in cell culture and biochemical applications. It is a structural component of epithelial cells and plays a role in the integrity and function of skin and hair. Keratin-10 is used in research settings to study cellular processes and protein interactions.

Automatically generated - may contain errors

Lab products found in correlation

Keratin 6, by Fortrea (3 mentions) Collagen 7, by Merck Group (2 mentions) Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (2 mentions) Loricrin, by Fortrea (2 mentions) Filaggrin, by Fortrea (2 mentions) Keratin 5, by Fortrea (2 mentions) Involucrin, by Merck Group (1 mentions) Cytochrome c, by BD (1 mentions) Occludin, by BD (1 mentions) Keratin 76, by Merck Group (1 mentions) Phospho histone h3, by Cell Signaling Technology (1 mentions) E cadherin, by Thermo Fisher Scientific (1 mentions) Proliferating cell nuclear antigen (pcna), by Santa Cruz Biotechnology (1 mentions) Claudin 1, by Abcam (1 mentions) Mouse anti β tubulin clone tub 2, by Merck Group (1 mentions) Ob cadherin, by Cell Signaling Technology (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) P cadherin, by Cell Signaling Technology (1 mentions) Mouse anti syndecan 1 clone b a38, by Abcam (1 mentions) Mouse anti ha 11 clone 16b12, by Fortrea (1 mentions)

Close spelling variants of this product

Rabbit anti-keratin 10 (5 citations) Keratin 10 (PRB-159P; (3 citations) Anti-Keratin 10 (PRB-159P) (2 citations)

9 protocols using keratin 10

Comprehensive Histological Analysis of Skin Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fixed skin tissues were embedded in paraffin and sectioned. The sections
were stained with hematoxylin & eosin, hematoxylin alone, or with
hematoxylin combined with the following antibodies: BrdU, total p53,
γ-H2AX [14 (link)]; Keratin-6 (rabbit
polyclonal, 1:500, Covance), Keratin-10 (rabbit polyclonal, 1:500, Covance), or
TRP2 (rabbit polyclonal, 1:500, Abcam). The tissues were then labeled with
secondary antibodies: Envision+ System HRP labeled polymer anti-rabbit HRP
(Dako) was used for the rabbit polyclonal antibodies. Melanin was stained with a
Fontana-Masson stain kit (Sigma-Aldrich Procedure No. HT200). Stained slides
were digitally scanned using a ScanScope CS (Aperio).

Lange S.S., Bhetawal S., Reh S., Powell K.L., Kusewitt D.F, & Wood R.D. (2018). DNA polymerase ζ deficiency causes impaired wound healing and stress-induced skin pigmentation. Life Science Alliance, 1(3), e201800048.

+ Open protocol

+ Expand

Immunofluorescence Microscopy of Cryosections

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Whole mount cryosections were prepared for immunofluorescence microscopy as previously described (Ridky et al., 2010 (link)). In short, slides were fixed in 4% paraformaldehyde or −20°C methanol, permeabilized as required and blocked with 10% horse serum/PBS, followed by incubation with primary antibodies and secondary antibodies conjugated to fluorophores. Slides were mounted with Prolong Gold Antifade Reagent with DAPI (Life Technologies, Grand Island, NY). The primary antibodies used in this study were keratin-5, keratin-10, loricrin, and filaggrin (Covance, Conshohocken, PA), Collagen VII (Millipore, Billerica, MA), involucrin (Sigma, St. Louis, MO), phospho-S6 (235/236), phospho-S6 (240/244), cleaved caspase-3, cleaved caspase-8, LC3A (Cell Signaling, Danvers, MA), p-PKC∂1 (Cell Signaling, Danvers, MA), TFAM (gift from C. Cameron, Penn State University), cytochrome c (BD Pharmingen, San Diego, CA), TRF2 (R & D Systems, Minneapolis, MN) and ki67 (ThermoScientific, Fremont, CA).

Monteleon C.L., Agnihotri T., Dahal A., Liu M., Rebecca V.W., Beatty G.L., Amavaradi R.K, & Ridky T.W. (2018). Lysosomes support the degradation, signaling, and mitochondrial metabolism necessary for human epidermal differentiation. The Journal of investigative dermatology, 138(9), 1945-1954.

+ Open protocol

+ Expand

Immunofluorescence Staining of Tight Junctions

Check if the same lab product or an alternative is used in the 5 most similar protocols

Staining for LacZ expression was performed as previously described [47] on frozen sections and counterstained with Nile Red. Immunofluorescence experiments were performed after citrate based antigen retrieval. Primary antibodies were ZO-1 (Invitrogen cat# 339100), Occludin (BD Transduction cat# 611090), Claudin-1 (ABCAM cat# ab15098), cytokeratin14 - LLOO2 (ABCAM cat # ab7800), keratin10 (Covance PRB-159P), keratin6 (Covance cat # PRB-169P), Ecadherin (Life Technologies, 13-1900), phospho-histone H3 (Cell Signalling, #9708), PCNA (Santa Cruz Biotechnology sc-9857), CLDN1 (Santa Cruz Biotechnology, sc-81796), Keratin 76 (Sigma-Aldrich HPA019696) and Keratin 76 (Sigma-Aldrich HPA019656), Filaggrin (FLG- Covance PRB-417P), FASN (Santa Cruz Biotechnology, sc-48357), and Melan-A (MEL-A, Santa Cruz Biotechnology, sc-20032). All secondary antibodies were AlexaFluor conjugated (Invitrogen).

DiTommaso T., Cottle D.L., Pearson H.B., Schlüter H., Kaur P., Humbert P.O, & Smyth I.M. (2014). Keratin 76 Is Required for Tight Junction Function and Maintenance of the Skin Barrier. PLoS Genetics, 10(10), e1004706.

+ Open protocol

+ Expand

Immunofluorescence Antibody Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Monoclonal antibodies used in this study included rabbit anti-αSMA antibody (clone E184; Abcam), mouse anti–α-actinin (clone BM75.2; Sigma-Aldrich), mouse anti-HA.11 (clone 16B12; Covance), mouse anti–β-tubulin (clone TUB 2.1; Sigma-Aldrich), mouse anti-paxillin (clone 5H11; EMD Millipore), mouse anti–syndecan-1 (clone B-A38; Abcam), rat anti–syndecan-4 (clone KY/8.2; BD), mouse anti-p120 catenin (clone 6H11; Invitrogen), rat anti–E-cadherin (clone DECMA-1; Abcam), and rabbit anti-NFAT3 (clone 23E6; Cell Signaling Technology). Polyclonal antibodies used were rabbit antibodies against syndecan-4 (Abcam and LSBio), OB-cadherin (Cell Signaling Technology), N-cadherin (Abcam), P-cadherin (Cell Signaling Technology), keratin-10 (Covance), TRPC4 (Sigma-Aldrich), and TRPC7 (Sigma-Aldrich and EMD Millipore). Secondary antibodies included goat anti–rabbit, goat anti–mouse, and rabbit anti–rat IgG HRP-conjugated antibodies from Dako. Donkey and goat anti–mouse, goat anti–rabbit, and donkey and goat anti–rat IgG conjugated to Alexa Fluor 488, 568, or 647 were obtained from Molecular Probes. DAPI and Alexa Fluor 568– and 647–conjugated phalloidin were obtained from Molecular Probes.

Gopal S., Søgaard P., Multhaupt H.A., Pataki C., Okina E., Xian X., Pedersen M.E., Stevens T., Griesbeck O., Park P.W., Pocock R, & Couchman J.R. (2015). Transmembrane proteoglycans control stretch-activated channels to set cytosolic calcium levels. The Journal of Cell Biology, 210(7), 1199-1211.

+ Open protocol

+ Expand

Protein Immunoblotting and Immunofluorescence for Epidermal Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein lysate from total epidermis was used for detection of Esrp1 and Esrp2. 10 μg of protein lysate was separated on Bis-Tris 4–20% Gradient SDS-PAGE gels (Invitrogen) and transferred to nitrocellulose membranes. Membranes were blocked in 5% Non-fat Milk in PBST then incubated overnight at 4°C in primary antibody using mouse monoclonal antibody to Esrp1 and Esrp2, 23A7, at 1:1000 dilution (Rockland Immunochemicals). Sheep anti-mIgG:HRP at 1:2500 (GE Healthcare) and ECL detection (Inivtrogen) by chemiluminescence. Loading control using anti-Beta Actin M2 (Sigma) at 1:10,000 and sheep anti-mIgG:HRP at 1:10,000 dilution. 7 μm sections from paraffin embedded dorsal skin was dewaxed and rehydrated according to traditional proceedures, and antigen retrieval was performed using antigen unmasking solution (Vector Laboratories). Primary antibodies for Keratin14 (AF 64) (Covance, Princeton, NJ), Keratin10 (Covance), Loricrin (AF 62) (Covance), FIaggrin (Covance), p63 (4A4) (Santa Cruz), β-catenin (15B8) (Sigma, St. Louis, MO), and Lef-1 (C18A7) (Cell Signalling) were used at 4° overnight, followed by fluorescent secondary antibodies Goat-anti-rIgG AlexaFlour 488 or Goat-anti-mIgG F(ab′)2 594 (Invitrogen). Images were taken using an Olympus BX43.

Bebee T.W., Park J.W., Sheridan K.I., Warzecha C.C., Cieply B.W., Rohacek A.M., Xing Y, & Carstens R.P. (2015). The splicing regulators Esrp1 and Esrp2 direct an epithelial splicing program essential for mammalian development. eLife, 4, e08954.

+ Open protocol

+ Expand

Immunostaining with Diverse Antibodies

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following antibodies were used for immunostainings. β-catenin (Sigma, sigmaaldrich.com), ABC (Millipore, Billerica, MA, Millipore. com), CD34 (BD Pharmingen, San Jose, CA, BDbioSciences.com), Keratin14 (Covance, Covance.com), Keratin 10 (Covance), AE13 (Abcam), Keratin17 (gift of P. Coulombe), pSmad 1/5/8 (Cell signaling, Boston, MA, cellsignal.com), NfatC1 (Santa Cruz Bio, Dallas, Tx, scbt. com), NFKB p50 (Santa Cruz Bio), and Caspase 3 (Cell Signaling). Sections were stained using standard immunohistochemical staining procedures.

Yucel G., Van Arnam J., Means P.C., Huntzicker E., Altindag B., Lara M.F., Yuan J., Kuo C, & Oro A.E. (2014). Partial Proteasome Inhibitors Induce Hair Follicle Growth by Stabilizing β-catenin. Stem cells (Dayton, Ohio), 32(1), 85-92.

+ Open protocol

+ Expand

Histological Analysis of Skin Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fixed skin tissues were embedded in paraffin and sectioned. The sections were stained with hematoxylin and eosin, hematoxylin alone, or with hematoxylin combined with the following antibodies: BrdU, total p53, and γ-H2AX (14 (link)); keratin-6 (rabbit polyclonal, 1:500; Covance); keratin-10 (rabbit polyclonal, 1:500; Covance); or TRP2 (rabbit polyclonal, 1:500; Abcam). The tissues were then labeled with secondary antibodies.EnVision+ System HRP labeled polymer; anti-rabbit HRP (Dako) was used for the rabbit polyclonal antibodies. Melanin was stained with a Fontana-Masson stain kit (Sigma-Aldrich procedure no.: HT200). Stained slides were digitally scanned using a ScanScope CS (Aperio).

+ Open protocol

+ Expand

Quantitative Immunofluorescence Analysis of Epidermal Differentiation

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Whole mount cryosections were prepared for immunofluorescence microscopy as previously described (Ridky et al., 2010 (link)). In short, slides were fixed in 4% paraformaldehyde or −20°C methanol, permeabilized as required and blocked with 10% horse serum/PBS, followed by incubation with primary antibodies and secondary antibodies conjugated to fluorophores. Slides were mounted with Prolong Gold Antifade Reagent with DAPI (Life Technologies, Grand Island, NY). The primary antibodies used in this study were collagen-VII (Millipore, Burlington, MA), ß1-integrin (Abcam, Cambridge, MA), ki67, desmoglein-3 (Thermo Fisher Scientific, Carlsbad, CA), HA, keratin-10, keratin-5, loricrin, and filaggrin (Covance, Dedham, MA). To quantify differentiation, the ratio of the area in pixels of keratin-10 positive epidermis to area in pixels of total epidermis was measured as a percentage in ImageJ. This analysis was based previously reported methods (Billings et al., 2015, Natale et al., 2018). Results are the mean of at least 3 technical replicates across at least three biologic replicates from individual donors (± s.d.). Significance was assessed by Welch’s t-test across the biological replicates.

Monteleon C.L., Lee I.Y, & Ridky T.W. (2019). Exophilin-5 supports lysosome-mediated trafficking required for epidermal differentiation. The Journal of investigative dermatology, 139(10), 2219-2222.e6.

+ Open protocol

+ Expand

Histological Analysis of Skin Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tissue sections (6 μm) were prepared and stained with hematoxylin and eosin (H&E) or Gomori’s trichrome containing methyl (aniline) blue for analysis of collagen I/III (Histopathology Core Facility, Rutgers University, Piscataway, NJ). For immunohistochemistry, tissue sections were deparaffinized, and blocked at room temperature with 1% goat or 1% horse serum for 2 hr and then incubated at room temperature for 30 min, or overnight at 4°C with primary rabbit affinity purified polyclonal antibodies to loricrin (1:400, Abcam, Cambridge, MA), keratin 10 or keratin 17 (1:200, Covance, Princeton, NJ), or with monoclonal antibodies to E-cadherin (1:300, Cell Signaling, Danvers, MA), or PCNA (1:300, Millipore, Burlington, MA) and mouse IgG (ProSci Inc., Poway, CA), or rabbit IgG (ProSci Inc.) as controls. After washing, tissue sections were incubated for 30 min with biotinylated goat anti-rabbit- or goat anti-mouse-secondary antibodies (Vector Labs, Burlingame, CA). Antibody binding was visualized using a DAB Peroxidase Substrate Kit (Vector Labs). Images of tissue sections were acquired at high resolution using an Olympus VS120 Virtual Microscopy System and analyzed using OlyVIA version 2.9 software (Center Valley, PA).

Laskin J.D., Wahler G., Croutch C.R., Sinko P.J., Laskin D.L., Heck D.E, & Joseph L.B. (2020). Skin remodeling and wound healing in the Gottingen minipig following exposure to sulfur mustard. Experimental and molecular pathology, 115, 104470.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!