Ki-67/MKI67 Antibody (NB500-170)

Images

 
Immunocytochemistry/ Immunofluorescence: Ki-67/MKI67 Antibody [NB500-170] - Ki67 Antibody [NB500-170] - Ki67 antibody was tested at 1:25 in HeLa cells with FITC (green). Nuclei and actin were counterstained with Dapi ...read more
Immunohistochemistry-Paraffin: Ki-67/MKI67 Antibody [NB500-170] - Ki67 Antibody [NB500-170] - Human tonsil.
Immunocytochemistry/ Immunofluorescence: Ki-67/MKI67 Antibody [NB500-170] - Ki67 Antibody [NB500-170] - Confocal immunofluorescent analysis of MCF7 cells using Ki67 antibody (NB500-170, 1:5). An Alexa Fluor ...read more
Western Blot: Ki-67/MKI67 Antibody [NB500-170] - Ki67 Antibody [NB500-170] - Western blot analysis of A431 (A) and Hek293 (B) cell lysate using Ki67 antibody (NB500-170) at 2 ug/ml.
Immunocytochemistry/ Immunofluorescence: Ki67 Antibody [NB500-170] - Ki67 antibody (1:100) was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, PoSpecies Glossary
Applications WB, ICC/IF, IHC, IHC-Fr, IHC-P
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.0 mg/ml

Ki-67/MKI67 Antibody Summary

Immunogen
A synthetic peptide made to an internal region of human Ki67 (within residues 1550-1700) [Swiss-Prot# P46013]
Localization
Nuclear
Marker
Proliferation Marker
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
MKI67
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1-2 ug/ml
  • Immunocytochemistry/Immunofluorescence 1:20-1:100
  • Immunohistochemistry 1:50-1:300
  • Immunohistochemistry-Frozen 1:50-1:300
  • Immunohistochemistry-Paraffin 1:50-1:300
Application Notes
This Ki67 antibody is useful for Immunohistochemistry on frozen and paraffin-embedded sections and Immunocytochemistry/Immunofluorescence. There have been mixed results in Western blot; however, NB500-170 has been used successfully Western Blog reported by a customer review. *Formalin fixed paraffin embedded tissue sections require high temperature antigen unmasking with 10 mM citrate buffer, pH 6.0 prior to immunostaining. This antibody will not work without optimal antigen retrieval. This is probably the most critical step. NOTE: We suggest an incubation period of 30 minutes at room temperature and to use DAB to stain the protein (immunofluorescence may give problems as the protein is nuclear).
Reviewed Applications
Read 5 Reviews rated 4.6
using
NB500-170 in the following applications:

Publications
Read Publications using
NB500-170 in the following applications:

Reactivity Notes

Human, mouse, rat (pubmed 22521325) and porcine.

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.05% Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

Alternate Names for Ki-67/MKI67 Antibody

  • antigen identified by monoclonal Ki-67
  • antigen KI-67
  • antiKi67
  • Ki67 ihc
  • Ki-67 ihc
  • Ki67 mouse
  • Ki-67 mouse
  • Ki67 western blot
  • Ki-67 western blot
  • Ki67
  • Ki-67
  • KIA
  • Marker Of Proliferation Ki-67
  • MIB-1
  • MKI67
  • PPP1R105
  • proliferation-related Ki-67 antigen
  • Protein Phosphatase 1
  • Regulatory Subunit 105
  • TSG126

Background

Originally discovered employing mouse monoclonal antibody against a nuclear antigen from Hodgkin's lymphoma-derived cell line, this non-histone protein was named Ki67 after researcher's location (Gerdes and colleagues), Ki for Kiel University in Germany and 67 referring to the clone number on the 96-well plate. It interacts with KIF15 as well as MKI67IP, and is suggested to be involved in cell cycle regulation. Ki67 is a large protein with expected molecular weight of about 395 kD and has a very complex localization pattern within the nucleus, one which changes during cell cycle progression. Its expression occurs specially during late G1, S, G2 and M phases of the cell cycle, while in cells undergoing G0 phase, Ki67 remains undetectable. Ki67 undergoes phosphorylation/dephosphorylation during mitosis, is susceptible to proteases and its structure implies that its expression is regulated by proteolytic pathways. Ki67 is associated with nucleolar DFC (dense fibrillary component) and its regulation appears to be tightly controlled (estimated half life is 60-90 min, regardless of the cell position in the cell cycle), presumably by precise synthesis and degradation systems involving proteasome, a protease complex. Due to its association with cell divison process, Ki-67 is routinely used as cellular proliferation marker of solid tumors as well as certain hematological malignancies, and a correlation has been demonstrated between Ki-67 index and the histopathological grade of cancers.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Applications: WB, ICC/IF, IHC, IHC-Fr, IHC-P

Publications for Ki-67/MKI67 Antibody (NB500-170)(59)

We have publications tested in 4 confirmed species: Human, Mouse, Rat, Porcine.

We have publications tested in 4 applications: ICC/IF, IHC, IHC-Fr, IHC-P.


Filter By Application
ICC/IF
(10)
IHC
(14)
IHC-Fr
(6)
IHC-P
(18)
All Applications
Filter By Species
Human
(20)
Mouse
(19)
Rat
(3)
Porcine
(2)
All Species
Showing Publications 1 - 10 of 59. Show All 59 Publications.
Publications using NB500-170 Applications Species
Tan Q, Joshua AM, Wang M et al. Up-regulation of autophagy is a mechanism of resistance to chemotherapy and can be inhibited by pantoprazole to increase drug sensitivity. Cancer Chemother. Pharmacol. May 1 2017 [PMID: 28378028] (Human) Human
Jayaraman S, Doucet M, Kominsky SL. Down-regulation of CITED2 attenuates breast tumor growth, vessel formation and TGF-B-induced expression of VEGFA. Oncotarget. Jan 24 2017 [PMID: 28008154] (Human) Human
Peng Y, Chen FF, Ge J et al. miR-429 Inhibits Differentiation and Promotes Proliferation in Porcine Preadipocytes. Int J Mol Sci. Dec 7 2016 [PMID: 27941616] (ICC/IF, Porcine) ICC/IF Porcine
Maeda A, BSc, Chen Y et al. In Vivo Imaging Reveals Significant Tumor Vascular Dysfunction and Increased Tumor HIF-1alpha Expression Induced by High Single-Dose Irradiation in a Pancreatic Tumor Model Int J Radiat Oncol Biol Phys Sep 14 2016 [PMID: 27816364] (IHC, Mouse) IHC Mouse
Zimmermann N, Thormann V, Hu B et al. Human isotype-dependent inhibitory antibody responses against Mycobacterium tuberculosis. EMBO Mol Med Nov 2 2016 [PMID: 27729388] (IHC-P, ICC/IF, Human) IHC-P, ICC/IF Human
Xiong J, Zhou MF, Wang YD et al. Protein Kinase D2 Protects against Acute Colitis Induced by Dextran Sulfate Sodium in Mice Sci Rep Sep 23 2016 [PMID: 27659202] (IHC, Human) IHC Human
Huang SW, Lien JC, Kuo SC, Huang TF. DDA suppresses angiogenesis and tumor growth of colorectal cancer in vivo through decreasing VEGFR2 signaling. Oncotarget. Sep 27 2016 [PMID: 27517319] (IHC, Mouse) IHC Mouse
Rock K, Joosse SA, Muller J et al. Chronic UVB-irradiation actuates perpetuated dermal matrix remodeling in female mice: Protective role of estrogen. Sci Rep. Jul 27 2016 [PMID: 27460287] (IHC, Mouse) IHC Mouse
Lohse I, Rasowski J, Cao P et al. Targeting hypoxic microenvironment of pancreatic xenografts with the hypoxia-activated prodrug TH-302. Oncotarget. Jun 7 2016 [PMID: 27248663] (Human) Human
de la Puente P, Muz B, Gilson RC et al. 3D tissue-engineered bone marrow as a novel model to study pathophysiology and drug resistance in multiple myeloma Biomaterials 2015 Sep 12 [PMID: 26402156] (IHC-P, Human)

Details:
This citation used the Alexa Fluor 488 version of this antibody.
IHC-P Human
Show All 59 Publications.

Reviews for Ki-67/MKI67 Antibody (NB500-170) (5) 4.65

Average Rating: 4.6
(Based on 5 reviews)
We have 5 reviews tested in 2 species: Human, Mouse.
We have 5 reviews tested in 4 application: ICC/IF, ICC, IHC-P, WB.

Reviews using NB500-170:
Filter by Applications
ICC/IF
(1)
ICC
(2)
IHC-P
(1)
WB
(1)
All Applications
Filter by Species
Human
(3)
Mouse
(2)
All Species
Images Ratings Applications Species Date Details
Immunocytochemistry/Immunofluorescence Ki-67/MKI67 NB500-170
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reviewed by:
Alexandra Ambrico
ICC/IF Mouse 10/11/2016
View

Summary

ApplicationImmunocytochemistry/Immunofluorescence
Sample Testedmouse prostate
SpeciesMouse
Immunocytochemistry Ki-67/MKI67 NB500-170
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reviewed by:
Natalie Erdmann
ICC Human 08/24/2015
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Summary

ApplicationImmunocytochemistry
Sample TestedMultiple myeloma cells lines LP-1 and JJN3 cytospins
SpeciesHuman
LotG-4

Blocking

Blocking Details10% donkey serum in PBS 1 hour at room temperature

Primary Anitbody

Dilution Ratio1/100 1ary antibody diluted in 10% donkey serum/PBS 3 hours at room temperature

Secondary Antibody

Secondary DescriptionDonkey F(ab")2 anti-rabbit IgG (H+L) Alexa Fluor 488
Secondary Manufacturer Cat#Jackson 711-546-152
Secondary Concentration1/1000

Details

Detection NotesZeiss AxioImager, FITC filter cube for 100 ms exposure
Fixation DetailsImmersion-fixed in 2% paraformaldehyde 10 minutes, 0.5% Triton X-100 5 minutes permeabilization at room temperature
Wash Description3X 5 minutes PBS after each step

Comments

CommentsWould use this reagent at 1/200 overnight at 4oC
 
reviewed by:
Anonymous
IHC-P Human 12/29/2014
View

Summary

ApplicationImmunohistochemistry-Paraffin
Sample TestedSee PMID 19582877
SpeciesHuman
Immunocytochemistry Ki-67/MKI67 NB500-170
Enlarge
reviewed by:
Anony- mous
ICC Human 06/09/2014
View

Summary

ApplicationImmunocytochemistry
SpeciesHuman
Special ApplicationsConfocal immunofluorescent analysis of MCF7 cells using Ki67 antibody (NB500-170, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).
FileView PDF
 
reviewed by:
Jian Zong
WB Mouse 09/30/2010
View

Summary

ApplicationWestern Blot
Sample TestedWhole cell lysate
SpeciesMouse

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for Ki-67/MKI67 Antibody (NB500-170). (Showing 1 - 2 of 2 FAQs).

  1. Could you please let us know if you have the antibody which may have cross-reactivity to hamster tissue in IHC(paraffin or frozen)? Also, please tell us reference information using your products to hamster tissue.
    • Thank you for your patience. The only information I was able to gain is that the protein has 61% homology with the chinese hamster. Our lab only recommends use if the % homology is 85% or above so I would not recommend our Ki67 antibodies for your customer. However if they are willing to still test this they would become eligable for our Innovators Reward Program in which they would gain a 50% refund and a 50% discount on a future purchase. All you would need to do is email us at innovators@novusbio.com. Here is the link to all of our Ki67 antibodies.
  2. What is the IF protocol which was used to obtain the IF image on the datasheet?
    • Our lab used our general ICC protocol to obtain the image seen on our website.

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB500-170AF405
Alexa Fluor 488 Labeled NB500-170AF488
Alexa Fluor 647 Labeled NB500-170AF647
Alexa Fluor 700 Labeled NB500-170AF700
Biotin Labeled NB500-170B
DyLight 405 Labeled NB500-170V
DyLight 488 Labeled NB500-170G
DyLight 550 Labeled NB500-170R
DyLight 650 Labeled NB500-170C
FITC Labeled NB500-170F

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Blogs on Ki-67/MKI67.

The relationship between Ki67 and HIF-1 in cancer
Ki67, also known as MKI67, is best known as the leading marker of cellular proliferation. Ki67 is regulated by a balance between synthesis and degradation, and often carries a very short half-life.  First discovered to be located to dividing cells,...  Read full blog post.

Ki67 - an established marker for labelling proliferating cells
Ki-67/MKI67 is an antigen which is expressed during G1, S, G2, and M phases of the cell cycle (mitotically active cells), but not during G0 phase (resting cells). It is a large protein with expected molecular weight of about 395 kDa, and it has a v...  Read full blog post.

Ki67 - A Crucial Cellular Proliferation Marker
The Ki67 antigen is a prototypic cell cycle-related protein expressed by proliferating cells in all phases of the active cell cycle (G1, S, G2 and M). It is a non-histone nuclear protein originally identified in a Hodgkin's lymphoma-derived cell line....  Read full blog post.

The Ki67 Antibody in Cell Marker Studies
The MK167, or Ki67 antibody recognizes a nuclear protein encoded by the MK167 gene. Ki167 is involved with RNA transcription and essential to cellular proliferation, being expressed by proliferating cells at all stages of the active cell cycle; it is...  Read full blog post.

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Recent Reviews

4.6
5
3
4
2
3
0
2
0
1
0

Alexandra Ambrico
10/11/2016
Application: ICC/IF
Species: Mouse

Natalie Erdmann
08/24/2015
Application: ICC
Species: Human

 
Anonymous
12/29/2014
Application: IHC-P
Species: Human

Bioinformatics

Gene Symbol MKI67
Entrez
Uniprot

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