NDC80 Antibody (9G3.23)

Images

 
Western Blot: NDC80 Antibody (9G3.23) [NB100-338] - HEC1 antibody [9G3.23] detects HEC1 protein by western blot analysis. Various whole cell extracts and HeLa nuclear extracts (30 ug) were separated by 7.5% SDS-PAGE, ...read more
Immunocytochemistry/ Immunofluorescence: NDC80 Antibody (9G3.23) [NB100-338] - HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: Hec1 stained by Hec1 antibody [9G3.23] diluted at 1:500. Red: alpha ...read more
Western Blot: NDC80 Antibody (9G3.23) [NB100-338] - WB analysis of NDC80 in U2OS Whole Cell Lysate. Verified customer review from 1DegreeBio.
Western Blot: NDC80 Antibody (9G3.23) [NB100-338] - Whole cell extract (30 ug) was separated by 7.5% SDS-PAGE, and the membrane was blotted with HEC1 antibody [9G3.23] diluted at 1:1000. The HRP-conjugated anti-mouse ...read more
Immunocytochemistry/ Immunofluorescence: NDC80 Antibody (9G3.23) [NB100-338] - Mouse anti-Hec1 and rabbit anti-Hec1 (phospho Ser165) antibodies were used to co-stain MCF10A cells. Insets show kinetochores of misaligned ...read more
Immunocytochemistry/ Immunofluorescence: NDC80 Antibody (9G3.23) [NB100-338] - Confocal immunofluorescence analysis (Olympus FV10i) of paraformaldehyde-fixed U2OS, using HEC1(NB100-338) antibody (Green) at 1:500 ...read more
Immunocytochemistry/ Immunofluorescence: NDC80 Antibody (9G3.23) [NB100-338] - IF analysis of NDC80 in fixed human cell lines of RPE1, H4, and 42MGBA. Image courtesy of anonymous customer product review.
Immunocytochemistry/ Immunofluorescence: NDC80 Antibody (9G3.23) [NB100-338] - IF analysis of NDC80 in PTK1 cells.Verified customer review from 1DegreeBio.

Product Details

Summary
Reactivity Hu, Rt, HaSpecies Glossary
Applications WB, Flow, ICC/IF, IHC, IHC-Fr, IP
Clone
9G3.23
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated

NDC80 Antibody (9G3.23) Summary

Immunogen
Human HEC1 protein consisting of amino acids 56-642.
Localization
Nuclear
Isotype
IgG2a
Clonality
Monoclonal
Host
Mouse
Gene
NDC80
Purity
Protein G 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 1:500-1:3000
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence 1:100-1:1000
  • Immunohistochemistry 1:10 - 1:500
  • Immunohistochemistry-Frozen 1:10 - 1:500
  • Immunoprecipitation 1-5 ug/ml
Theoretical MW
80 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Positive Control
HeLa Lysate (NB800-PC1)
Reviewed Applications
Read 5 Reviews rated 4.6
using
NB100-338 in the following applications:

Publications
Read Publications using
NB100-338 in the following applications:

  • 33 publications
  • WB
    4 publications

Reactivity Notes

Kangaroo rat.

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
No Preservative
Purity
Protein G purified

Alternate Names for NDC80 Antibody (9G3.23)

  • HEC1KNTC2
  • HECRetinoblastoma-associated protein HEC
  • Highly expressed in cancer protein
  • highly expressed in cancer, rich in leucine heptad repeats (yeast)
  • highly expressed in cancer, rich in leucine heptad repeats
  • hsHec1
  • hsNDC80
  • kinetochore associated 2
  • Kinetochore protein Hec1
  • kinetochore protein NDC80 homolog
  • Kinetochore-associated protein 2
  • NDC80 homolog, kinetochore complex component (S. cerevisiae)
  • TID3

Background

Human HEC1 is a coiled-coil protein that has been demonstrated to have a role in M-phase progression. It is essential for proper chromosome segregation. Its depletion results in the impairment of chromosome congression and results in spindle checkpoint activation.

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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Publications for NDC80 Antibody (NB100-338)(42)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 2 applications: ICC/IF, WB.


Filter By Application
ICC/IF
(33)
WB
(4)
All Applications
Filter By Species
Human
(11)
All Species
Showing Publications 1 - 10 of 42. Show All 42 Publications.
Publications using NB100-338 Applications Species
Raab M, Sanhaji M, Matthess Y et al. PLK1 has tumor-suppressive potential in APC-truncated colon cancer cells. Nat Commun. Mar 16 2018 [PMID: 29549256] (ICC/IF, Human) ICC/IF Human
Dumitru AMG, Rusin SF, Clark AEM et al. Cyclin A/Cdk1 modulates Plk1 activity in prometaphase to regulate kinetochore-microtubule attachment stability Elife. 2017 Nov 20 [PMID: 29154753] (ICC/IF, Human)

Details:
HEC1 IF antibody used at a dilution of 1:500
ICC/IF Human
Niikura Y, Kitagawa R, Ogi H, Kitagawa K. SGT1-HSP90 complex is required for CENP-A deposition at centromeres Cell Cycle 2017 Aug 17 [PMID: 28816574] (Human) Human
Tauchman EC, Boehm FJ, DeLuca JG. Stable kinetochore-microtubule attachment is sufficient to silence the spindle assembly checkpoint in human cells. Nat Commun. 2015 Dec 01 [PMID: 26620470] (WB, ICC/IF, Human) WB, ICC/IF Human
Kleyman M, Kabeche L, Compton DA. STAG2 promotes error correction in mitosis by regulating kinetochore-microtubule attachments. J. Cell. Sci. 2014 Jul 29 [PMID: 25074805] (ICC/IF, Human)

Details:
Outer kinetochore component Hec1/NDC80 antibody used in ICC-IF on HCT116 and RPE1 cells for measuring centromere stretch (Fig. 1D, supplementary material Fig. S2A)
ICC/IF Human
Sivakumar S, Daum JR, Tipton AR, Gorbsky GJ. The spindle and kinetochore-associated (Ska) complex enhances binding of the anaphase-promoting complex/cyclosome (APC/C) to chromosomes and promotes mitotic exit. Mol. Biol. Cell 2014 Mar 1 [PMID: 24403607] (WB, Human) WB Human
Yao R, Kondoh Y, Natsume Y et al. A small compound targeting TACC3 revealed its different spatiotemporal contributions for spindle assembly in cancer cells. Oncogene. 2013 Sep 30 [PMID: 24077290] (WB, Human) WB Human
Kabeche L, Compton DA. Cyclin A regulates kinetochore microtubules to promote faithful chromosome segregation. Nature. 2013 Oct 3 [PMID: 24013174] (WB, Human) WB Human
Maier B, Kirsch M, Anderhub S et al. The novel actin/focal adhesion-associated protein MISP is involved in mitotic spindle positioning in human cells. Cell Cycle 2013 May 1 [PMID: 23574715] (ICC/IF, Human) ICC/IF Human
De Antoni A, Maffini S, Knapp S et al. A small-molecule inhibitor of Haspin alters the kinetochore functions of Aurora B. J Cell Biol 2012 Oct [PMID: 23071153] (ICC/IF) ICC/IF
Show All 42 Publications.

Reviews for NDC80 Antibody (NB100-338) (5) 4.65

Average Rating: 4.6
(Based on 5 reviews)
We have 5 reviews tested in 2 species: Human, Other.

Reviews using NB100-338:
Filter by Applications
IF
(4)
WB
(1)
All Applications
Filter by Species
Human
(4)
Other
(1)
All Species
Images Ratings Applications Species Date Details
Immunofluorescence NDC80 NB100-338
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reviewed by:
Anonymous
IF Human 04/21/2016
View

Summary

ApplicationImmunofluorescence
Sample TestedRPE1 cells
SpeciesHuman

Blocking

Blocking DetailsBlocked in PBS-BSA (1% BSA) for 10 minutes

Primary Anitbody

Dilution RatioAntibody was diluted in PBS-BSA (1%BSA) at 1:1000 and incubated for 1hr at room temperature

Secondary Antibody

Secondary DescriptionDonkey anti-rabit Alexafluor 488
Secondary Manufacturer Cat#Jackson Immuno Laboratory
Secondary ConcentrationAntibody was diulted at 1:1000 in PBS-BSA (1%BSA) for 1hr at room temperature

Details

Detection NotesNikon TiE was used for imaging. Single focal plane image at 60X
Fixation DetailsFixed with 3.5% paraformaldehyde for 15 minuted. Permeabilized with PBS+0.5% TritonX100 for 5 minutes. Washed in PBS for 5mins
Wash Description1 Wash in PBS for 5 minutes
Western Blot NDC80 NB100-338
Enlarge
reviewed by:
Andrew Cosgrove
WB Human 11/06/2013
View

Summary

ApplicationWestern Blot
Sample TestedU2OS whole cell lysate
SpeciesHuman

Blocking

Blocking DetailsBlock with TBST + 5% Milk for 1 hr

Primary Anitbody

Dilution Ratio1:2000 in TBST + 2% Milk for 1hr

Secondary Antibody

Secondary ConcentrationIn TBST + 2% Milk for 1hr
Immunofluorescence NDC80 NB100-338
Enlarge
reviewed by:
Andrew Cosgrove
IF Other 11/06/2013
View

Summary

ApplicationImmunofluorescence
Sample TestedPTK1 cells
SpeciesOther
CommentsRed is mouse anti-Hec1 with TxRed secondary

Blocking

Blocking DetailsBSA

Primary Anitbody

Dilution Ratio1:1000 in TBS-BSA + 0.1% TritonX for 1 hr

Details

Fixation DetailsFixed in 3.5% paraformaldehyde for 15 mins
Wash DescriptionWash with TBS-BSA for 5 mins

Comments

CommentsRed is mouse anti-Hec1 with TxRed secondary
 
reviewed by:
Anonymous
IF Human 10/04/2013
View

Summary

ApplicationImmunofluorescence
SpeciesHuman
Pub Med ID24013174
FileView PDF
Immunofluorescence NDC80 NB100-338
Enlarge
reviewed by:
Duane Compton
IF Human 06/06/2012
View

Summary

ApplicationImmunofluorescence
Sample TestedFixed cell lines (RPE1, H4, 42MGBA)
SpeciesHuman

Blocking

Blocking DetailsTBS-BSA +.5% triton --> 10min rocking, TBS-BSA --> 10min rocking

Primary Anitbody

Dilution Ratio1:1000 in TBS-BSA, 4C overnight

Secondary Antibody

Secondary DescriptionALEXA Cy5 donkey anti-mouse IgG
Secondary Concentration1:500

Details

Fixation Details3.5% paraformaldehyde, 10 minutes room-temp
Wash DescriptionTBS-BSA 10min rocking

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 NDC80 Antibody (NB100-338) (0)

There are no specific FAQs related to this product. Read our general customer & technical service FAQs.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB100-338AF405
Alexa Fluor 488 Labeled NB100-338AF488
Alexa Fluor 647 Labeled NB100-338AF647
Alexa Fluor 700 Labeled NB100-338AF700
Allophycocyanin Labeled NB100-338APC
DyLight 350 Labeled NB100-338UV
DyLight 405 Labeled NB100-338V
DyLight 488 Labeled NB100-338G
DyLight 680 Labeled NB100-338FR
DyLight 755 Labeled NB100-338IR
PE Labeled NB100-338PE
PerCP Labeled NB100-338PCP

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Recent Reviews

4.6
5
3
4
2
3
0
2
0
1
0

Anonymous
04/21/2016
Application: IF
Species: Human

Andrew Cosgrove
11/06/2013
Application: WB
Species: Human

Andrew Cosgrove
11/06/2013
Application: IF
Species: Other

Bioinformatics

Gene Symbol NDC80
Entrez
Uniprot