discontinued

BM4538 Cytokeratin (Hair Cortex) antibody

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0.1 ml / €450.00

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Mouse anti Human, Mouse Cytokeratin (Hair Cortex) AE13

BM4538

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  • BM4538

Product Description for Cytokeratin (Hair Cortex)

Mouse anti Human, Mouse Cytokeratin (Hair Cortex) AE13.
Presentation: Purified
Product is tested for Flow Cytometry, Western blot / Immunoblot, Paraffin Sections.

Properties for Cytokeratin (Hair Cortex)

Product Category Primary Antibodies
Quantity 0.1 ml
Presentation Purified
Reactivity Hu, Ms
Applications F, P, WB
Clonality Monoclonal
Clone AE13
Host Mouse
Isotype IgG
Molecular weight 44-46 kDa (Predicted)
Shipping to Worldwide
PDF datasheet View Datasheet
Manufacturer Acris Antibodies GmbH
Material safety datasheet MSDS for Monoclonal Antibodies (de)

Datasheet Extract

Immunogen
Immunogen:
Human hair keratins.
Application Immunblotting: 1/1000-1/3000. Detects a band of approximately 44 kDa.
Flow Cytometry: 1/20-1/50.
Immunohistochemistry on Frozen Sections (Ref.1-7).
Immunohistochemistry on Paraffin Sections.
Background The keratins are intermediate filament proteins responsible for the structural integrity of epithelial cells and are subdivided into cytokeratins and hair keratins. Most of the type I cytokeratins consist of acidic proteins which are arranged in pairs of heterotypic keratin chains and are clustered in a region on chromosome 17q21.2.
Concentration 1 mg/ml
Protocols Immunofluorescence Protocol - Formaldehyde Fixation
Collect cells from T.c.unit and remove media from petri dish using suction.
Wash with 1x PBS and remove.
Incubate cells in pre-warm (37°C) Para-Formaldehyde for 12 minutes at room temperature on an orbital shaker.
Remove PFA and incubate in 0.5% Triton X-IOO in 1x PBS for 5 minutes at room temperature.
Prepare blocking reagent, this is also the antibody diluent.
Wash cells 2x with 1x PBS at room temperature, for 4 minutes/wash on an orbital shaker.
Block with 1 % NCS and 1x PBS for 30 minutes at room temperature.
Prepare primary antibodies (50µl/coverslip) and moist staining chambers.
Wash cells 2x with lx PBS at room temperature and air dry briefly.
Incubate with primary antibody for 1 hr at room temperature in the dark in staining chambers. During this time prepare the secondary antibody.
Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)
Incubate with secondary antibody for 1 hour at room temperature in the dark in staining chambers.
Wash cells 5x with 1x PBS.
Mount in Dapi.
Solutions (prepare fresh the same day of staining):
1x Phosphate buffered saline.
Blocking reagent: 1% NCS in 1x PBS (use fresh l0x PBS).
Fixation Solution: 3.5% Para formaldehyde.
1.75g PFA in 20 ml d.H20 plus 5 drops 1M NaOH. Stir on a hot plate at 50-60°C until dissolved. Add 4 drops IN HCI and check pH indicator strip. PH should be 7.4. Complete volume with d.H20 to 25ml and add 25ml 2xPBS. Check pH before adding to cover slips.
Immunofluorescence protocol - Methanol/Acetone Fixation
Collect cells from T.C.unit and remove media from petri dish using suction.
Wash with 1x PBS and remove.
Fix cells with cold methanol: acetone 1:1 for 10 minutes on ice.
Prepare blocking reagent, this is also the diluent for the antibodies.
Remove fixative and wash cells 3x with Ix PBS at RT, for 4 minutes/wash on orbital shaker.
Block with 1% NCS and Ix PBS for 30 minutes at RT.
Prepare primary antibodies (50µl/coverslip) and moist staining chambers.
Wash cells 2x with 1 x PBS at RT and air dry for approximately 7 minutes.
Incubate with primary antibody for 1 hr at RT in the dark in staining chambers. During this time prepare secondary antibody.
Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)
Incubate with secondary antibody for 1 hr at R T in the dark in staining chambers.
Wash cells 5x with 1x PBS.
Mount in Dapi.
Solutions (prepare fresh the same day of staining):
1x Phosphate buffered saline.
Blocking reagent: 1% NCS in 1x PBS (use fresh 10x PBS).
Fixation solution: methanol:acetone 1: 1 ice cold.
Western Blotting Protocol
Transfer gel to PDVF or nitrocellulose membrane
Place membrane in plastic tray in blocking buffer for one hour with agitation
Rinse in wash buffer
Incubate in wash buffer plus primary antibody for one hour
Wash 6 X 5 minutes with wash buffer
Incubate in wash buffer plus secondary antibody for one hour
Wash 6X 5 minutes with wash buffer
Detect (e.g. ECL, Amersham according to manufacturers instructions)
Wash buffer: PBS + 0.1% Tween 20
Blocking buffer: Wash buffer + 5% dried milk powder
The concentration of antibodies used depends on each antibody, the amount of antigen and the detection method used. Generally, dilution is in the range of a few hundred times dilution to a few thousand times dilution, but usually has to be determined empirically.
General Readings
  1. Chen D, Jarrell A, Guo C, Lang R, Atit R. Dermal β-catenin activity in response to epidermal Wnt ligands is required for fibroblast proliferation and hair follicle initiation. Development. 2012 Apr;139(8):1522-33. doi: 10.1242/dev.076463. PubMed PMID: 22434869. (Free PMC Article available, 7 images available)
  2. Larouche D, Cuffley K, Paquet C, Germain L. Tissue-engineered skin preserving the potential of epithelial cells to differentiate into hair after grafting. Tissue Eng Part A. 2011 Mar;17(5-6):819-30. doi: 10.1089/ten.TEA.2010.0403. Epub 2010 Dec 14. PubMed PMID: 20973750.
  3. Youssef KK, Van Keymeulen A, Lapouge G, Beck B, Michaux C, Achouri Y, et al. Identification of the cell lineage at the origin of basal cell carcinoma. Nat Cell Biol. 2010 Mar;12(3):299-305. doi: 10.1038/ncb2031. Epub 2010 Feb 14. PubMed PMID: 20154679.
  4. Van Keymeulen A, Mascre G, Youseff KK, Harel I, Michaux C, De Geest N, et al. Epidermal progenitors give rise to Merkel cells during embryonic development and adult homeostasis. J Cell Biol. 2009 Oct 5;187(1):91-100. doi: 10.1083/jcb.200907080. Epub 2009 Sep 28. PubMed PMID: 19786578. (Free PMC Article available, 5 images available)
  5. List K, Szabo R, Molinolo A, Nielsen BS, Bugge TH. Delineation of matriptase protein expression by enzymatic gene trapping suggests diverging roles in barrier function, hair formation, and squamous cell carcinogenesis. Am J Pathol. 2006 May;168(5):1513-25. PubMed PMID: 16651618. (Free PMC Article available, 7 images available)
  6. Kajikawa M, Baba T, Tomaru U, Watanabe Y, Koganei S, Tsuji-Kawahara S, et al. MHC class I-like MILL molecules are beta2-microglobulin-associated, GPI-anchored glycoproteins that do not require TAP for cell surface expression. J Immunol. 2006 Sep 1;177(5):3108-15. PubMed PMID: 16920948.
  7. Sun Y, Strizzi L, Raafat A, Hirota M, Bianco C, Feigenbaum L, et al. Overexpression of human Cripto-1 in transgenic mice delays mammary gland development and differentiation and induces mammary tumorigenesis. Am J Pathol. 2005 Aug;167(2):585-97. PubMed PMID: 16049342. (Free PMC Article available, 7 images available)
  8. Suzuki K, Yamaguchi Y, Villacorte M, Mihara K, Akiyama M, Shimizu H, et al. Embryonic hair follicle fate change by augmented beta-catenin through Shh and Bmp signaling. Development. 2009 Feb;136(3):367-72. doi: 10.1242/dev.021295. PubMed PMID: 19141668.
  9. Lynch MH, O'Guin WM, Hardy C, Mak L, Sun TT. Acidic and basic hair/nail ("hard") keratins: their colocalization in upper cortical and cuticle cells of the human hair follicle and their relationship to "soft" keratins. J Cell Biol. 1986 Dec;103(6 Pt 2):2593-606. PubMed PMID: 2432071. (Free PMC Article available)
  10. Dhouailly D, Xu C, Manabe M, Schermer A, Sun TT. Expression of hair-related keratins in a soft epithelium: subpopulations of human and mouse dorsal tongue keratinocytes express keratin markers for hair-, skin- and esophageal-types of differentiation. Exp Cell Res. 1989 Mar;181(1):141-58. PubMed PMID: 2465162.
Storage Store undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.
Avoid repeated freezing and thawing.
Shelf life: one year from despatch.
Format
Purification:
Protein G Chromatography
Buffer System:
PBS, pH 7.2
Preservatives:
0.09% Sodium Azide
State:
Liquid purified IgG fraction
Purified
Specificity
Specificity:
This antibody recognises Acidic 44-46 kDa hair cortex keratins.
AE13 is an excellent marker for hair and nail differentiation.
Species:
Human and Mouse.
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