Invitrogen

HEC1 Monoclonal Antibody (9G3.23)

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

9 Published Figures

9 References

View all (21) HEC1 antibodies

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Datasheet

Protocols

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HEC1 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of HEC1 (kinetochore, red) in a BHP 2-7 thyroid cancer anaphase cell cultured in complete medium on a glass slide. Cells were fixed with 4% phosphate-buffered paraformaldehyde for 30 min at 37C, permeabilized with 100% ice-cold methanol for 20 min. and blocked with PBS/1% BSA/0.5% Tween 20 at 37C for 30 min. Cells were stained with anti-HEC1, clone 9G3, mouse IgG2a monoclonal antibody (Product # MA1-23308) at a dilution of 1:200 in PBS/1% BSA/0.1% Tween 20 for 60 min at 37C., and then incubated with goat anti-mouse IgG2a AF594 (Product # A-21135) at a dilution of 1:200 for 60 min. at 37C (panel a: red). Alpha tubulin (panel b: green) was stained simultaneously with anti alfa-tubulin, clone DM1A, mouse IgG1 (Product # 14-4502-82) followed by goat anti-mouse IgG1 AF488 (1:200, Cat # A-21121). DNA was stained with Hoechst33342 (panel c: blue). Panels d shows a merged image. Images were taken with a 40x immersion oil objective (for more information: Corver et al., Endocr Relat Cancer. 2018 Jan;25(1):83-97). Data courtesy of Antibody Data Exchange Program.

HEC1 Antibody in Western Blot (WB) — Western blot was performed using Anti-HEC1 Monoclonal Antibody (9G3.23) (Product # MA1-23308) and a 74 kDa band corresponding to HEC1 was observed in Hela, U-2 OS, Jurkat, NCCIT and K-562. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), U-2 OS (Lane 2), Jurkat (Lane 3), NCCIT (Lane 4) and K-562 (Lane 5) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

HEC1 Antibody in Western Blot, Immunocytochemistry (WB, ICC/IF) — Figure 4. Reduced outer kinetochore size of PSCs in mitosis. ( a ) Scheme representing the architecture and interactions of different proteins that comprise the human centromere and kinetochore. ( b ) Representative immunofluorescence images from differentiated (RPE and fibroblasts derived from donor no. 1 and donor no. 2-Fibr D no. 1 and Fibr D no. 2) and pluripotent stem cells (human embryonic stem cell line H9-hESC and iPSCs reprogrammed from Fibr D no. 1 and Fibr D no. 2-iPSC D no. 1 and iPSC D no. 2) for CENP-E and Hec1. ( c ) Quantitation of centromeric CENP-E and Hec1. Mean levels of fluorescence per nuclei was measured (CENP-E) or average centromere intensities were determined using automatic centromere recognition and quantification (Hec1). The average and standard error of the mean of one (Hec1) or three (CENP-E) independent experiments are shown. ( d ) Human ESCs, RPE, iPSCs and the fibroblasts they were reprogrammed from, were harvested and processed for SDS-PAGE and immunobloting. CENP-E and Hec1 levels were assessed with specific antibodies. GAPDH (Hec1) or tubulin (CENP-E) was used as a loading control. CENP-E and CENP-A (in figure 2 ) were detected in the same gel shown using different channels. ( e ) Quantitation of WB bands. Average and standard error of the mean of three independent experiments are shown. Protein levels were normalized to GAPDH or tubulin. Scale bar = 2 um.

HEC1 Antibody in Immunohistochemistry (IHC) — Fig. 5 SENP6 is required for maintaining the integrity of the CCAN independent of proteolysis. a Schematic representing the architecture and interactions of different protein complexes in the human centromere and kinetochore. b Centromeric levels of different CCAN proteins following auxin-mediated depletion of SENP6 (24 h). Cells were counterstained with CENP-B to mark centromeres. Bars, 10 um. c Centromeric levels of KMN (KNL1-MIS12-NDC80) members following auxin-mediated depletion of SENP6. Bars, 10 um. d Automated centromere recognition and quantification of b and c . Fluorescence intensities of indicated proteins were normalized to the mean of the control 'no auxin' condition in each experiment and plotted as a bar graph for auxin treated '+' or no auxin '-' condition. Three replicate experiments were performed. Bars represent SEM. Parametric two-tailed Student's t test were performed to calculate statistical significance. ** p < 0.01, *** p < 0.001. e Immunoblot showing the total levels of CENP-A and CENP-C proteins following 48 h treatment with auxin to deplete AID-SENP6. Extracts from control '-' or auxin treated '+' cells were separated by SDS-PAGE and immunoblotted with anti-CENP-A (detecting both SNAP tagged and endogenous CENP-A) or anti-CENP-C antibody. Tubulin was used as loading control. Source data are provided as a Source Data file.

HEC1 Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HeLa cells were transfected with HEC1 siRNA and reduction of signal was observed in Western Blot using MALT1 Monoclonal Antibody (Product # MA1-23308) {KD}

Product Details

MA1-23308

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:3,000	View 2 publications 2 publications
Immunohistochemistry (IHC)	-	View 1 publication 1 publication
Immunohistochemistry (Frozen) (IHC (F))	Assay-dependent	-
Immunocytochemistry (ICC/IF)	1:100-1:1,000	View 6 publications 6 publications
Flow Cytometry (Flow)	Assay-dependent	-
Immunoprecipitation (IP)	1-5 µg/mL	-
in situ PLA (PLA)	Assay-dependent	-

Product Specifications
Species Reactivity	Hamster, Human, Mouse, Non-human primate, Pig, Rodent
Published species	Human
Host/Isotype	Mouse / IgG2a
Class	Monoclonal
Type	Antibody
Clone	9G3.23
Immunogen	Partial human HEC1 protein consisting of amino acids 56-642. View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.88 mg/mL
Purification	Protein A
Storage buffer	PBS
Contains	no preservative
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2149871

Product Specific Information

Positive control of HELA WCE suggested.

Target Information

Hec1 is one of several proteins involved in spindle checkpoint signaling. This surveillance mechanism assures correct segregation of chromosomes during cell division by detecting unaligned chromosomes and causing prometaphase arrest until the proper bipolar attachment of chromosomes is achieved.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

CENP-A overexpression promotes aneuploidy with karyotypic heterogeneity.

The Journal of cell biology

Shrestha RL,Rossi A,Wangsa D,Hogan AK,Zaldana KS,Suva E,Chung YJ,Sanders CL,Difilippantonio S,Karpova TS,Karim B,Foltz DR,Fachinetti D,Aplan PD,Ried T,Basrai MA

Published figure using HEC1 monoclonal antibody (Product # MA1-23308) in Immunocytochemistry

Mon Apr 05 00:00:00 EDT 2021

Induction of spontaneous human neocentromere formation and long-term maturation.

The Journal of cell biology

Murillo-Pineda M,Valente LP,Dumont M,Mata JF,Fachinetti D,Jansen LET

MA1-23308 was used in Immunocytochemistry to report a chromosome engineering system for neocentromere formation in human cells and characterize the first experimentally induced human neocentromere at a naive locus.

Mon Mar 01 00:00:00 EST 2021

Induction of spontaneous human neocentromere formation and long-term maturation.

The Journal of cell biology

Murillo-Pineda M,Valente LP,Dumont M,Mata JF,Fachinetti D,Jansen LET

Mon Mar 01 00:00:00 EST 2021

chTOG is a conserved mitotic error correction factor.

eLife

Herman JA,Miller MP,Biggins S

MA1-23308 was used in Western Blot, Immunocytochemistry to demonstrate there is likely a conserved function for the TOG protein family.

Wed Dec 30 00:00:00 EST 2020

chTOG is a conserved mitotic error correction factor.

eLife

Herman JA,Miller MP,Biggins S

MA1-23308 was used in Western Blot, Immunocytochemistry to demonstrate there is likely a conserved function for the TOG protein family.

Wed Dec 30 00:00:00 EST 2020

chTOG is a conserved mitotic error correction factor.

eLife

Herman JA,Miller MP,Biggins S

MA1-23308 was used in Western Blot, Immunocytochemistry to demonstrate there is likely a conserved function for the TOG protein family.

Wed Dec 30 00:00:00 EST 2020

chTOG is a conserved mitotic error correction factor.

eLife

Herman JA,Miller MP,Biggins S

MA1-23308 was used in Western Blot, Immunocytochemistry to demonstrate there is likely a conserved function for the TOG protein family.

Wed Dec 30 00:00:00 EST 2020

A Biosensor for the Mitotic Kinase MPS1 Reveals Spatiotemporal Activity Dynamics and Regulation.

Current biology : CB

Kuijt TEF,Lambers MLA,Weterings S,Ponsioen B,Bolhaqueiro ACF,Staijen DHM,Kops GJPL

MA1-23308 was used in Immunocytochemistry to develop MPS1sen, a sensitive and specific FRET-based biosensor for MPS1 activity o understand when and where MPS1 activation occurs and how MPS1 signaling is modulated during mitosis.

Mon Oct 05 00:00:00 EDT 2020

Reprogramming of human cells to pluripotency induces CENP-A chromatin depletion.

Open biology

Milagre I,Pereira C,Oliveira RA,Jansen LET

MA1-23308 was used in Immunocytochemistry, Western Blot to explore the composition of the centromere and kinetochore in human embryonic and induced PSCs.

Thu Oct 01 00:00:00 EDT 2020

Reprogramming of human cells to pluripotency induces CENP-A chromatin depletion.

Open biology

Milagre I,Pereira C,Oliveira RA,Jansen LET

MA1-23308 was used in Immunocytochemistry, Western Blot to explore the composition of the centromere and kinetochore in human embryonic and induced PSCs.

Thu Oct 01 00:00:00 EDT 2020

Reprogramming of human cells to pluripotency induces CENP-A chromatin depletion.

Open biology

Milagre I,Pereira C,Oliveira RA,Jansen LET

MA1-23308 was used in Immunocytochemistry, Western Blot to explore the composition of the centromere and kinetochore in human embryonic and induced PSCs.

Thu Oct 01 00:00:00 EDT 2020

Genetic screening identifies a SUMO protease dynamically maintaining centromeric chromatin.

Nature communications

Mitra S,Bodor DL,David AF,Abdul-Zani I,Mata JF,Neumann B,Reither S,Tischer C,Jansen LET

Published figure using HEC1 monoclonal antibody (Product # MA1-23308) in Immunohistochemistry

Fri Jan 24 00:00:00 EST 2020

The quantitative architecture of centromeric chromatin.

eLife

Bodor DL,Mata JF,Sergeev M,David AF,Salimian KJ,Panchenko T,Cleveland DW,Black BE,Shah JV,Jansen LE

MA1-23308 was used in immunocytochemistry to study the localization of CENP-A in the cell and at centromeres.

Tue Jul 15 00:00:00 EDT 2014

Bioinformatics

Protein Aliases: HEC-1; HEC2; HEC4; Highly expressed in cancer protein; highly expressed in cancer, rich in leucine heptad repeats; HsHec1; kinetochore associated 2; Kinetochore protein Hec1; Kinetochore protein NDC80 homolog; Kinetochore-associated protein 2; NDC80 homolog, kinetochore complex component; NDC80 kinetochore complex component; NDC80 kinetochore complex component homolog; putative kinetochore protein NDC80 like protein; Retinoblastoma-associated protein HEC

Gene Aliases: 2610020P18Rik; AV139730; HEC; HEC1; HsHec1; hsNDC80; KNTC2; NDC80; TID3

UniProt ID: (Human) O14777, (Mouse) Q9D0F1

Entrez Gene ID: (Human) 10403, (Pig) 100626545, (Chinese hamster) 100751668, (Mouse) 67052

Molecular Function: transcription cofactor

Function(s)

mitotic sister chromatid segregation establishment of mitotic spindle orientation mitotic spindle organization chromosome segregation sister chromatid cohesion mitotic nuclear division attachment of spindle microtubules to kinetochore cell division attachment of mitotic spindle microtubules to kinetochore cell cycle

Process(es)

structural constituent of cytoskeleton protein binding identical protein binding

HEC1 Monoclonal Antibody (9G3.23)

Product Details

MA1-23308

Western Blot (WB)

Immunohistochemistry (IHC)

Immunohistochemistry (Frozen) (IHC (F))

Immunocytochemistry (ICC/IF)

Flow Cytometry (Flow)

Immunoprecipitation (IP)

in situ PLA (PLA)

Species Reactivity

Published species

Host/Isotype

Class

Type

Clone

Immunogen

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

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