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BD Horizon™ PE-CF594 Mouse Anti-Human Cytokeratin
Clone CAM5.2 (RUO)
Flow cytometric analysis of cytokeratin expression in human SK-BR-3 cells. Cells from the human SK-BR-3 (Breast adenocarcinoma, ATCC HTB-30) cell line were incubated with 1X BD FACS™ Lysing Solution (Cat. No. 349202), permeabilized with BD FACS™ Permeabilizing Solution 2 (Cat. No. 340973/347692), and washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656). The cells were then stained with either BD Horizon™ PE-CF594 Mouse Anti-Human Cytokeratin antibodies (Cat. No. 563615; solid line histogram) or BD Horizon™ PE-CF594 Mouse IgG2a, κ Isotype Control (Cat. No. 562306; dashed line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of cytokeratin expression in human SK-BR-3 cells. Cells from the human SK-BR-3 (Breast adenocarcinoma, ATCC HTB-30) cell line were incubated with 1X BD FACS™ Lysing Solution (Cat. No. 349202), permeabilized with BD FACS™ Permeabilizing Solution 2 (Cat. No. 340973/347692), and washed with BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656). The cells were then stained with either BD Horizon™ PE-CF594 Mouse Anti-Human Cytokeratin antibodies (Cat. No. 563615; solid line histogram) or BD Horizon™ PE-CF594 Mouse IgG2a, κ Isotype Control (Cat. No. 562306; dashed line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Horizon™
Cytokeratin-7/-8; CK-7/CK-8;
Human (QC Testing)
Mouse BALB/c IgG2a
Human HT29 colorectal carcinoma cell line
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2738319
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 with BD Horizon™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were removed.
Recommended Assay Procedures
Note: The CAM5.2 antibody can also be used to stain cells that have been fixed and permeabilized using BD Cytofix™ Fixation Buffer (Cat. No. 554655) and BD Perm/Wash™ Buffer (Cat. No. 554723).
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.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- CF™ is a trademark of Biotium, Inc.
- When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
- Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
- 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
Lysing Solution 10X Concentrate IVD
Size
100 ML
Cat No.
349202
PE-CF594 Mouse IgG2a, κ Isotype Control RUO
Size
50 µg
Cat No.
563484
PE-CF594 Mouse IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
562306
Permeabilizing Solution 2 RUO
Size
500 Tests
Cat No.
340973
Permeabilizing Solution 2 RUO (GMP)
Size
200 Tests
Cat No.
347692
563615 Rev. 1
Antibody Details
CAM5.2
The CAM5.2 monoclonal antibody specifically recognizes cytokeratin having a primary reactivity with human keratin proteins that correspond to Moll's peptides #7 (48 kDa) and #8 (52 kDa). Cytokeratin 7 and 8 are type II cytoskeletal keratins. These cytoskeletal proteins provide structural integrity for epithelial cells and may serve other functions as well. They are expressed in epithelia cells that comprise normal human tissues. Although these cytokeratins are not normally expressed in stratified squamous epithelium, they may be expressed in some squamous cell carcinomas. The CAM 5.2 antibody stains most epithelial-derived tissue, including liver, renal tubular epithelium, and hepatocellular and renal cell carcinomas.
This antibody is conjugated to BD Horizon™ PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red®. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red® yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red® (eg 610/20-nm filter).
563615 Rev. 1
Format Details
PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and Yellow-Green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and Yellow-Green (561-nm) lasers and the acceptor dye can be excited by the Green (532-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE-CF594
496 nm, 566 nm
615 nm
563615 Rev.1
Citations & References
View product citations for antibody "563615" on CiteAb
Development References (6)
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Battifora H. Diagnostic uses of antibodies to keratins: A review and immunohistochemical comparison of seven monoclonal and three polyclonal antibodies. In: Fenoglio-Preiser C, Wolff M, Rilke F, ed. Progress in Surgical Pathology. 1988:1-15.
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Cooper D, Schermer A, Sun TT. Biology of disease classification of human epithelia and their neoplasms using monoclonal antibodies to keratins: strategies, applications and limitations. Lab Invest. 1985; 52:243-256. (Clone-specific). View Reference
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Leader M, Patel J, Makin C, Henry K. An analysis of the sensitivity and specificity of the cytokeratin marker CAM 5.2 for epithelial tumors: results of a study of 203 sarcomas, 50 carcinomas, and 28 malignant melanomas. Histopathology. 1986; 10:1315-1324. (Clone-specific: Immunohistochemistry). View Reference
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Makin C, Bobrow L, Bodmer W. Monoclonal antibody to cytokeratin for use in routine histopathology. J Clin Pathol. 1984; 37(9):975-983. (Immunogen). View Reference
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Moll R, Franke W, Schiller D, Geiger B, Krepler R. The catalog of human cytokeratins: patterns of expression in normal epithelia, tumors, and cultured cells. Cell. 1982; 31:11-24. (Clone-specific). View Reference
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Smedts F, Ramaekers F, Robben H, et al. Changing patterns of keratin expression during progression of cervical intraepithelial neoplasia. Am J Pathol. 1990; 136(3):657-668. (Clone-specific: Immunohistochemistry, Western blot). View Reference
563615 Rev. 1
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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.