Flow cytometry and Immunoflourescent analysis of Cytokeratin 14, 15, 16 and 19 in human A431 cells. (Left Panel) Human A431 (Human epithelial carcinoma; ATCC CRL-1555) cell line were fixed with a 1% solution of paraformaldehyde and permeabilized with 0.1% Triton™ X-100. The cells were washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) and then stained with either Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No. 557732/557783; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human Cytokeratin 14, 15, 16 and 19 antibody (Cat. No. 563648; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact A431 cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System. (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 Alexa Fluor® 647 Mouse Anti-Human Cytokeratin 14, 15, 16 and 19 monoclonal antibody (Cat. No. 563648; pseudo-colored red). The counter-staining of cell nuclei was with BD Pharmingen™ DAPI Solution (pseudo-colored blue)(Cat. No.564907). 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) Human A431 (Human epithelial carcinoma; ATCC CRL-1555) cell line were fixed with a 1% solution of paraformaldehyde and permeabilized with 0.1% Triton™ X-100. The cells were washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) and then stained with either Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No. 557732/557783; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human Cytokeratin 14, 15, 16 and 19 antibody (Cat. No. 563648; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact A431 cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System. (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 Alexa Fluor® 647 Mouse Anti-Human Cytokeratin 14, 15, 16 and 19 monoclonal antibody (Cat. No. 563648; pseudo-colored red). The counter-staining of cell nuclei was with BD Pharmingen™ DAPI Solution (pseudo-colored blue)(Cat. No.564907). 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
BD Pharmingen™
KRT14, KRT15, KRT16, KRT19; CK14, CK15, CK16, CK19; K14, K15, K16, K19
Human (QC Testing)
Mouse IgG1, κ
Intracellular staining (flow cytometry) (Routinely Tested), Immunofluorescence (Tested During Development)
5 µl
AB_2738347
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Triton is a trademark of the Dow Chemical Company.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
Perm Buffer III RUO
Size
125 mL
Cat No.
558050
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557732
Alexa Fluor® 647 Mouse IgG1 κ Isotype control RUO
Size
50 Tests
Cat No.
557783
563648 Rev. 3
Antibody Details
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.
563648 Rev. 3
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
563648 Rev.3
Citations & References
Development References (2)
-
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
-
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
563648 Rev. 3
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.
