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Safety Data Sheet (SDS) FITC Anti-mouse Ki-67 Antibody     Product Data Sheet (PDF)    
FITC Anti-mouse Ki-67 Antibody
3862045 25 µg $110.00       
3862050 100 µg $280.00       
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 FITC under optimal conditions. The solution is free of unconjugated FITC and unconjugated antibody.
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 ≤0.5 µg per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.

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) FITC (filled histogram) or rat IgG2a, κ FITC 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.