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Purified Mouse Anti-Human Cytokeratin 14, 15, 16 and 19
Purified Mouse Anti-Human Cytokeratin 14, 15, 16 and 19

Flow cytometry and Immunoflourescent analysis of Cytokeratin 14, 15, 16 and 19 in human A431 cells.

     (Left Panel) A431 cells were fixed with 1% paraformaldehyde, permeabilized with 0.1% Triton™ X-100 buffer, stained with second step reagent with FITC Goat Anti-Mouse IgG/IgM (Cat. No. 555988), and analyzed by flow cytometry.

     (Right Panel) Human A431 cells (Human epithelial carcinoma; ATCC CRL-1555) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with 0.1% Triton™ X-100 buffer, and stained with purified Mouse Anti-Human Cytokeratin 14, 15, 16 and 19 monoclonal antibody (Cat. No. 550951; pseudo-colored green) at 5μg/mL. The second step reagent was Alexa Fluor® 488 Goat Anti-Mouse Ig (Life Technologies) and the counter-staining of cell nuclei was with BD Pharmingen™ DAPI Solution (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ software. Permeabilization with BD Perm/Wash™ (Cat No. 554723) or BD Phosflow™ Perm Buffer III (Cat. No. 558050) is also suitable for use with this antibody.

Flow cytometry and Immunoflourescent analysis of Cytokeratin 14, 15, 16 and 19 in human A431 cells.

     (Left Panel) A431 cells were fixed with 1% paraformaldehyde, permeabilized with 0.1% Triton™ X-100 buffer, stained with second step reagent with FITC Goat Anti-Mouse IgG/IgM (Cat. No. 555988), and analyzed by flow cytometry.

     (Right Panel) Human A431 cells (Human epithelial carcinoma; ATCC CRL-1555) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with 0.1% Triton™ X-100 buffer, and stained with purified Mouse Anti-Human Cytokeratin 14, 15, 16 and 19 monoclonal antibody (Cat. No. 550951; pseudo-colored green) at 5μg/mL. The second step reagent was Alexa Fluor® 488 Goat Anti-Mouse Ig (Life Technologies) and the counter-staining of cell nuclei was with BD Pharmingen™ DAPI Solution (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ software. Permeabilization with BD Perm/Wash™ (Cat No. 554723) or BD Phosflow™ Perm Buffer III (Cat. No. 558050) is also suitable for use with this antibody.

Product Details
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BD Pharmingen™
Human (QC Testing)
Mouse IgG1, κ
Intracellular staining (flow cytometry) (Routinely Tested), Immunofluorescence (Tested During Development)
0.5 mg/ml
AB_393974
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. Triton is a trademark of the Dow Chemical Company.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
550951 Rev. 7
Antibody Details
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KA4

The KA4 monoclonal antibody specifically binds to cytokeratins 14, 15, 16 and 19 present in the cytoplasm of a wide variety of human simple epithelium. Human epithelium has been shown to contain cytoplasmic filaments of the cytokeratin type. There have been 19 different polypeptides identified and they demonstrate a characteristic pattern for each type of epithelium. This antibody is useful for intracellular staining and flow cytometric analysis, Western blot and immunohistochemical staining of both, acetone-fixed frozen tissue sections and formalin-fixed paraffin embedded tissue sections with citrate pretreatment.

550951 Rev. 7
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
550951 Rev.7
Citations & References
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Development References (2)

  1. Nagle RB, Lucas DO, McDaniel KM, Clark VA, Schmalzel GM. Paget's cells. New evidence linking mammary and extramammary Paget cells to a common cell phenotype. Am J Clin Pathol. 1985; 83(4):431-438. (Biology). View Reference
  2. Nagle RB, Moll R, Weidauer H, Nemetschek H, Franke WW. Different patterns of cytokeratin expression in the normal epithelia of the upper respiratory tract. Differentiation. 1985; 30(2):130-140. (Biology). View Reference
550951 Rev. 7

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.