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Purified 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
1_Ki-67_PE_3_100410
Resting (dashed line) or PHA-activated human peripheral blood lymphocytes (day-3, solid line) fixed and permeabilized with 70% ethanol, then intracellularly stained with Ki-67 PE.
  • 1_Ki-67_PE_3_100410
    Resting (dashed line) or PHA-activated human peripheral blood lymphocytes (day-3, solid line) fixed and permeabilized with 70% ethanol, then intracellularly stained with Ki-67 PE.
  • 2_Ki-67_Purified_Ki-67_Antibody_ICC_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 purified Ki-67 (clone Ki-67) (red) in blocking buffer overnight at 4°C and followed by DyLight™ 594 anti-mouse IgG and 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.
  • 3_Ki-67_PURE_Ki-67_Antibody_WB_121217
    Total lysates (15 µg protein) from 293T control (Ctrl) and Ki-67 CRISPR/Cas9 knock out (Ki-67 KO) cells were resolved by electrophoresis (4-20% Tris-glycine gel), transferred to nitrocellulose, and probed with 1:1000 (0.5 µg/ml) purified anti-Ki-67 antibody, clone Ki-67. Proteins were visualized using chemiluminescence detection by incubation with HRP Goat anti-Mouse secondary antibody (Cat. No. 405306, 1:3000 dilution). Direct-Blot™ HRP anti-β-actin antibody was used as a loading control (Cat. No. 643807, 1:8000 dilution).
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350501 25 µg 65 CHF
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350502 100 µg 147 CHF
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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.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

ICFC - Quality tested
CyTOF®, ICC, WB - Verified
IHC-F - Reported in the literature, not verified in house
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. For Western blotting, the suggested use of this reagent is 0.5 µg per ml (1:1000). It is recommended that the reagent be titrated for optimal performance for each application.

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

(PubMed link indicates BioLegend citation)
  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. Ferreira JCC, et al. 2022. Int J Mol Sci. 24: . PubMed
  2. Qin XY, et al. 2023. Cell Death Dis. 14:358. PubMed
  3. Santos R, et al. 2017. Nat Commun. . 10.1038/s41467-017-01760-5. PubMed
  4. Mann ER, et al. 2020. Sci Immunol. :5. PubMed
  5. Guha P, et al. 2013. Proc Natl Acad Sci U S A. 110:5052. PubMed
  6. Wang L, et al. 2022. Oncol Lett. 23:62. PubMed
  7. Junghof J, et al. 2022. NPJ Regen Med. 7:14. PubMed
  8. O'Boyle KC, et al. 2020. Methods Mol Biol. 2111:1. PubMed
  9. Bruno TC, et al. 2017. Cancer Immunol Res. 0.831944444. PubMed
  10. Agrawal N, et al. 2018. Front Immunol. 2.053472222. PubMed
  11. Xie S, et al. 2020. Ann Transl Med. 1.384027778. PubMed
  12. Eccles JD, et al. 2020. Cell Rep. 30:351. PubMed
  13. Tailor D, et al. 2021. Cell Chemical Biology. . PubMed
  14. Zhou H, et al. 2017. Leukemia. 31:2065. PubMed
  15. Xiong Y, et al. 2019. Onco Targets Ther. 12:993. PubMed
  16. Vining KH, et al. 2018. Adv Mater. 30:4. PubMed
  17. Wang Z et al. 2019. Br J Pharmacol. 176(17):3390-3406 . PubMed
RRID
AB_10662385 (BioLegend Cat. No. 350501)
AB_10662385 (BioLegend Cat. No. 350502)

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: 12.12.2017

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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