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Brilliant Violet 421™ 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
Ki-67_BV421_1_040311
3-day PHA-stimulated (top) or non-stimulated (bottom) human peripheral blood lymphocytes were fixed with 70% cold ethanol, then stained with Ki-67 Brilliant Violet 421™ (filled histogram) or mouse IgG1 Brilliant Violet 421™ isotype control (open histogram).
  • Ki-67_BV421_1_040311
    3-day PHA-stimulated (top) or non-stimulated (bottom) human peripheral blood lymphocytes were fixed with 70% cold ethanol, then stained with Ki-67 Brilliant Violet 421™ (filled histogram) or mouse IgG1 Brilliant Violet 421™ isotype control (open histogram).
  • Ki-67_BV421_2_040311
  • Ki-67_BV421_Ki-67_Antibody_3_081319
    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 2.5 µg/ml of Ki-67 (clone Ki-67) Brilliant Violet 421™ (blue) in blocking buffer overnight at 4°C and followed by Alexa Fluor® 488 Phalloidin (green) staining for 20 minutes. Nuclei were counterstained with DRAQ5 and are shown in red. The image was captured with 40X objective.
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350505 25 tests 168€
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350506 100 tests 344€
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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 Brilliant Violet 421™ 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.

Brilliant Violet 421™ excites at 405 nm and emits at 421 nm. The standard bandpass filter 450/50 nm is recommended for detection. Brilliant Violet 421™ is a trademark of Sirigen Group Ltd.


Learn more about Brilliant Violet™.

This product is subject to proprietary rights of Sirigen Inc. and is made and sold under license from Sirigen Inc. The purchase of this product conveys to the buyer a non-transferable right to use the purchased product for research purposes only. This product may not be resold or incorporated in any manner into another product for resale. Any use for therapeutics or diagnostics is strictly prohibited. This product is covered by U.S. Patent(s), pending patent applications and foreign equivalents.
Excitation Laser
Violet Laser (405 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.

Additional Product Notes

View more applications data using this product in multi-color fluorescence microscopy and intracellular flow cytometry in our Scientific Poster Library.

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. Leeansyah E, et al. 2015. PLoS Pathog. 11: 1005072. PubMed
  2. Vishnoi M, et al. 2018. Cancer Res. 78:5349. PubMed
  3. Taylor HE, et al. 2020. Cell Rep. 31:107810. PubMed
  4. Siriwon N, et al. 2018. Cancer Immunol Res. 6:812. PubMed
  5. Izmirly AM, et al. 2022. PLoS Pathog. 18:e1009903. PubMed
  6. Ackerley CG, et al. 2022. Front Immunol. 13:972170. PubMed
  7. Chierico L, et al. 2017. PLoS One. 12(2):e0171815. PubMed
  8. Cillo AR, et al. 2021. Cell Rep Med. 2:100476. PubMed
  9. Fu Y, et al. 2021. Cell Reports. 36(2):109344. PubMed
  10. Sprouse ML, et al. 2019. Int J Mol Sci. 20:8. PubMed
RRID
AB_10896915 (BioLegend Cat. No. 350505)
AB_2563860 (BioLegend Cat. No. 350506)

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

Related FAQs

What is the F/P ratio range of our BV421™ format antibody reagents?

It is lot-specific. On average it ranges between 2-4.

Go To Top Version: 5    Revision Date: 08-13-2019

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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Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

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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