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Alexa Fluor® 647 anti-human Ki-67 Antibody

Pricing & Availability
Clone
Ki-67 (See other available formats)
Regulatory Status
RUO
Other Names
Antigen Ki-67
Isotype
Mouse IgG1, κ
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Product Citations
publications
A_Ki-67_AF647_062911.jpg
PHA-stimulated (3 days) human peripheral blood lymphocytes were fixed and permeabilized with 70% ethanol, and then stained with Ki-67 Alexa Fluor® 647 (filled histogram) or mouse IgG1 κ Alexa Fluor® 647 isotype control (open histogram).
  • A_Ki-67_AF647_062911.jpg
    PHA-stimulated (3 days) human peripheral blood lymphocytes were fixed and permeabilized with 70% ethanol, and then stained with Ki-67 Alexa Fluor® 647 (filled histogram) or mouse IgG1 κ Alexa Fluor® 647 isotype control (open histogram).
  • B_Ki-67_AF647_Ki-647_Antibody_IF_032218
    HeLa cells were fixed with 1% paraformaldehyde (PFA) for 10 minutes, permeabilized with 0.5% Triton X-100 for 10 minutes, and blocked with 5% FBS for 30 minutes. The cells were then intracellularly stained with 5 µg/ml of Ki-67 (clone Ki-67) Alexa Fluor® 647 (red) in blocking buffer overnight at 4°C and followed by Alexa Fluor® 488 Phalloidin (green) staining for 20 minutes. Nuclei were counterstained with DAPI and are shown in blue. The image was captured with 40X objective.
Compare all formats See Alexa Fluor® 647 spectral data
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350509 25 tests 109€
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350510 100 tests 259€
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Description

Antigen Ki-67 is a nuclear protein expressed as two isoforms with molecular weights of 395 and 345 kD. Both isoforms contain one forkhead-associated domain and 16 concatenated "Ki-67 repeats," each containing the epitope recognized by the mAb Ki-67. The antigen Ki-67 interacts with Hklp2, hNIFK, and chromobox protein homolog 1, 3, and 5. Ki-67 is required for cell proliferation and its expression is restricted to the phases G1, S, G2, and M of the cell cycle. This characteristic makes Ki-67 an excellent marker for proliferating cells and is commonly used as one of the prognostic factors in cancer studies. Ki-67 has also been used to study myocyte proliferation after myocardial infarction as well as lymphocyte proliferation during infection, and has been used in neurons of patients with different neuropathologies.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human
Reported Reactivity
Cow
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Nuclei of the Hodgkin lymphoma cell line L428
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 647 under optimal conditions.
Concentration
Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
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
ICC - Verified
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 5 µl per million cells in 100 µl staining volume or 5 µl per 100 µl of whole blood. For immunocytochemistry, a concentration of 5.0 µg/ml is recommented. 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.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Red Laser (633 nm)
Application Notes

Additional reported applications (for the relevant formats) include: immunohistochemical staining of frozen tissue sections1, Western blotting3, and immunofluorescence microscopy4.

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 BioLegend 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 BioLegend Cell Staining Buffer and then resuspend in 0.5 ml cell staining buffer for flow cytometric analysis.

Application References
  1. Gerdes J, et al. 1983. Int. J. Cancer 31:13. (IHC)
  2. Gerdes J, et al. 1984. J. Immunol. 133:1710. (ICFC)
  3. Schluter C, et al. 1993 J. Cell Biol. 123:513. (IHC, WB)
  4. Bading H, et al. 1989 Exp. Cell. Res. 185:50. (IF)
  5. Guha P, et al. 2013. PNAS. 110:5052. PubMed
Product Citations
  1. Joshi SK, et al. 2021. Cancer Cell. . PubMed
  2. Trabanelli S, et al. 2017. Nat Commun. 10.1038/s41467-017-00678-2. PubMed
  3. Pei S et al. 2018. Cell stem cell. 23(1):86-100 . PubMed
  4. Ong SM, et al. 2018. Cell Death Dis. 9:266. PubMed
  5. Lindesmith LC, et al. 2020. Cell Mol Gastroenterol Hepatol. 0.586805556. PubMed
  6. Lin JR et al. 2018. eLife. 7 pii: e31657. PubMed
  7. Schulz E, et al. 2021. Toxins (Basel). 13:. PubMed
  8. Yokomizo-Nakano T, et al. 2020. Cancer Res. 80:2523. PubMed
  9. Watanabe N, et al. 2017. Oncoimmunology. 5:e1253656. PubMed
RRID
AB_10900810 (BioLegend Cat. No. 350509)
AB_10900810 (BioLegend Cat. No. 350510)

Antigen Details

Structure
Two isoforms with molecular weights of 395 and 345 kD, one forkhead-associated domain, 16 concatenated Ki-67 repeats, located in nucleus
Distribution

Expressed in the phases G1, S, G2, and M of the cell cycle

Function
Required for cell proliferation
Interaction
Chromobox protein homolog 1, 3 and 5, Hklp2, and hNIFK
Biology Area
Cell Biology, Cell Cycle/DNA Replication, DNA Repair/Replication
Molecular Family
Nuclear Markers
Antigen References

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

Gene ID
4288 View all products for this Gene ID
UniProt
View information about Ki-67 on UniProt.org
Go To Top Version: 6    Revision Date: 06.27.2014

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

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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