Cytokeratin 5 Antibody

Western Blot: Cytokeratin 5 Antibody [NBP2-61931] - Total protein from human A431 cells and rat PC12 cells was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-Cytokeratin 5 in 5% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.

Immunocytochemistry/Immunofluorescence: Cytokeratin 5 Antibody [NBP2-61931] - A431 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.1% Triton X-100. The cells were incubated with anti-Cytokeratin 5 at 5 ug/ml overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: Cytokeratin 5 Antibody [NBP2-61931] - Analysis of a FFPE tissue section of human skin using 1:1000 dilution of rabbit anti-Cytokeratin 5 antibody. The staining was developed using HRP labeled anti-rabbit secondary antibody and DAB reagent, and nuclei of cells were counter-stained with hematoxylin. This Cytokeratin 5 antibody generated a specific membrane cytoplasmic staining in the epithelial cells of the skin tissue. The signal was highest in the keratinocytes while dermis was largely negative for this Cytokeratin 5 (KRT5).

Flow Cytometry: Cytokeratin 5 Antibody [NBP2-61931] - An intracellular stain was performed on A431 cells with Cytokeratin 5 Antibody NBP2-61931AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 10 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

• Reactivity: Hu, Rt
• Applications: WB, Flow, ICC/IF, IHC, IHC-P
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

Cytokeratin 5 Antibody Summary

• Immunogen: Partial recombinant human Cytokeratin 5 protein (amino acids 19-78) [Uniprot: P13647]
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: KRT5
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/ml
  • Flow Cytometry 2-10 ug/million cells
  • Immunocytochemistry/Immunofluorescence 5 ug/ml
  • Immunohistochemistry 1:1000
  • Immunohistochemistry-Paraffin 1:1000

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for Cytokeratin 5 Antibody

• 58 kDa cytokeratin
• CK5
• CK-5
• cytokeratin-5
• DDD
• EBS2
• epidermolysis bullosa simplex 2 Dowling-Meara/Kobner/Weber-Cockayne types
• K5
• keratin 5 (epidermolysis bullosa simplex, Dowling-Meara/Kobner/Weber-Cockaynetypes)
• keratin 5
• keratin, type II cytoskeletal 5
• Keratin-5
• KRT5A
• Type-II keratin Kb5

Background

Keratins are a family of structurally related proteins that form the intermediate filament cytoskeleton in epithelial cells. The 58-kD keratin CK-5 is highly similar to other type II keratins and less similar to type I keratins and other intermediate filament proteins. The 58-kD keratin is regulated by retinoids in several tissues and is one of four keratins abundantly expressed in epidermal keratinocytes, where it may be important in maintaining structural integrity of the integument (1). Keratin 5 (CK-5) mRNA and protein are shown to be expressed in normal mammary epithelial cells in culture and are absent from tumor-derived cell lines. This makes CK-5 an important marker in the tumorigenic process, distinguishing normal from tumor cells, and decreased CK-5 expression correlates with tumorigenic progression (2). Dowling-Degos disease (DDD) is an autosomal dominant genodermatosis characterized by progressive and disfiguring reticulate hyperpigmentation of the flexures. Loss of function of CK-5 suggests a crucial role for keratins in the organization of cell adhesion, melanosome uptake, organelle transport, and nuclear anchorage (3).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: KRT5