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Safety Data Sheet (SDS) Alexa Fluor 647 Anti-mouse Ki-67 Antibody     Product Data Sheet (PDF)    
Alexa Fluor® 647 Anti-mouse Ki-67 Antibody
3862035 25 µg $120.00       
3862040 100 µg $320.00       
Clone: 16A8
Isotype: Rat IgG2a, κ
Reactivity: Mouse, Human
Immunogen: E. coli expressed partial mouse Ki-67 recombinant protein, 1816-2163 aa.
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation: The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 647 under optimal conditions.
Concentration: 0.5 mg/ml
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application:

ICFC - Quality tested
IF - Reported in the literature

Application Notes:

Ki-67 Staining Protocol:

1. Prepare 70% ethanol and chill at -20°C.
2. Prepare target cells of interest and wash 2X with PBS by centrifuge at 350xg for 5 minutes.
3. Discard supernatant and loosen the cell pellet by vortexing.
4. Add 3 ml cold 70% ethanol drop by drop to the cell pellet while vortexing.
5. Continue vortexing for 30 seconds and then incubate at -20°C for 1 hour.
6. Wash 3X with Cell Staining Buffer and then resuspend the cells at the concentration of 0.5-10 x 106/ml.
7. Mix 100 µl cell suspension with proper fluorochrome-conjugated Ki-67 antibody and incubate at room temperature in the dark for 30 minutes.
8. Wash 2X with Cell Staining Buffer and then resuspend in 0.5 ml cell staining buffer for flow cytometric analysis.

Recommended Usage:

Each lot of this antibody is quality control tested by our Ki-67 staining protocol below. For flow cytometric staining, the suggested use of this reagent is ≤0.25 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633 nm / 635 nm.

Application References:

1. Medina-Reyes EI, et al. 2015. Environ Res. 136:424. PubMed
2. Guillaumond F, et al. 2015. PNAS. 112:2473. PubMed
3. Sharma SK, et al. 2015. J Immunol. 194:5529. PubMed
4. Rodero MP, et al. 2014. J. Invest. Dermatol. 7:1991-7. PubMed

Con A-stimulated (3 days) BALB/c

Con A-stimulated (3 days) BALB/c mouse splenocytes were fixed and permeabilized with 70% ethanol, then stained with Ki-67 (clone 16A8) Alexa Fluor® 647 (filled histogram) or rat IgG2a, κ Alexa Fluor® 647 isotype control (open histogram).



Description:

The nuclear protein Ki-67 was first identified by the monoclonal antibody Ki-67, which was generated by immunizing mice with nuclei of the L428 Hodgkin lymphoma cell line. Ki-67 protein plays an essential role in ribosomal RNA transcription and cell proliferation. Expression of Ki-67 occurs during G1, S, G2, and M phase, while in G0 phase the Ki-67 protein is not detectable. Ki-67 is strongly expressed in proliferating cells and has been reported as a prognostic marker in various tumors.

Other Names: KIA, proliferation-related Ki-67 antigen
Structure: 325 kD protein containing a forkhead-associated domain (FHA) and 13 tandem repeats
Distribution: Nucleus and chromosome
Function: Required for cell cycle progression and proliferation
Interaction: Interacts with KIF15; binds to MKI67IP through FHA domain
Antigen References:

1. Starborg M, et al. 1996. J. Cell. Sci. 109:143.
2. Byeon IJ, et al. 2005. Nat. Struct. Mol. Biol. 12:987.
3. Yerushalmi R, et al. 2010. Lancet. Oncol. 11:174.
4. Beltrami AP, et al. 2001. N. Engl. J. Med. 344:1750.
5. Sachsenberg N, et al. 1998. J. Exp. Med. 187:1295.
6. Nagy Z, et al. 1997. Acta. Neuropathol. 93:294.


This product is provided under an agreement between Life Technologies Corporation and Sony Biotechnology Inc. and the manufacture, use, sale or import of this product may be subject to one or more U.S. patents, pending applications, and corresponding non-U.S. equivalents, owned by and/or licensed to Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research and/or development conducted by the buyer (whether the buyer is an academic or for profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to (1) use of the product or its components in manufacturing, (2) use of the product or its components for therapeutic, diagnostic or prophylactic purposes, or (3) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Molecular Probes Detection Technologies, Attn: Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.