Chromosome 10 Open Reading Frame 2 (C10ORF2) (Middle Region) antibody

Details for Product No. ABIN2775251
Antigen
  • ATXN8
  • IOSCA
  • MTDPS7
  • PEO
  • PEO1
  • PEOA3
  • SANDO
  • SCA8
  • TWINL
  • C6H10orf2
  • D19Ertd626e
  • Twinl
  • twinkle mtDNA helicase L homeolog
  • twinkle mtDNA helicase
  • twnk.L
  • TWNK
  • Twnk
Epitope
Middle Region
13
9
8
4
3
2
1
1
1
1
Reactivity
Cow (Bovine), Dog (Canine), Guinea Pig, Horse (Equine), Human, Mouse (Murine), Rabbit, Rat (Rattus), Zebrafish (Danio rerio)
73
33
31
2
2
2
1
1
1
1
Host
Rabbit
72
2
Clonality
Polyclonal
Conjugate
Un-conjugated
6
4
4
3
3
3
3
3
3
2
2
2
1
1
1
1
1
Application
Western Blotting (WB)
45
23
23
13
6
5
2
1
1
1
Options
'Independent Validation' Badge
Antigen PEO1 antibody - middle region
Lot Number QC6322
Method validated Western Blotting
Positive Control HeLa cells, HeLa cells overexpressing PEO1
Negative Control rho0 HeLa cells
Notes Passed, regarding sensitivity and specificity. The antibody detects an antigen at the expected apparent molecular weight of its antigen.
Primary Antibody ABIN2775251
Secondary Antibody Goat-anti rabbit IgG (H+L), HRP-linked (Invitrogen, A-16104)
Protocol
  • Cell lysates were propared from HeLa cells, HeLa cells overexpressing PEO1, and rho0 HeLa cells.
  • 75µg total protein or each sample were separated on a denaturing 8% SDS-PAGE gel (Laemmli 1970).
  • Immunoblot onto supported nitrocellulose membrane (Protran, GE Healthcare No. 1060005, 0.2µm) (Towbin et al., 1979).
  • Blocking of the membrane in 3% skim milk in TBST (50mM Tris-HCl, pH 7.4, 150mM NaCl, 0.1% Tween 20) for 1h at room temperature.
  • Incubation with primary antibody ABIN2775251 diluted 1:1000 in 3% BSA in TBST at 4°C overnight.
  • Washing in TBST for 3x 5min.
  • Incubation with goat-anti rabbit IgG (H+L), HRP-linked (Invitrogen, A-16104) diluted 1:15000 in 3% BSA in TBST for 1h at room temperature.
  • Washing in TBST for 5x 10min.
  • Chemiluminescence detection with luminol solution (250µg/ml Na-Luminol, 0,01% H2O2, 11µg/ml p-hydroxy-coumaric acid in 100mM Tris pH 8.5) and image capture on Kodak ECL film.
Experimental Notes PEO1 runs usually at 95-100 kDa, although the theoretical size is 72 kDa, often it is also visible as dimer.
Validation Images
Western Blotting validation image for anti-Chromosome 10 Open Reading Frame 2 (C10ORF2) (Middle Region) antibody (ABIN2775251) 75µg total protein of HeLa cells overexpressing PEO1 (lane 1), HeLa cells (lane 2), a...
'Independent Validation' Badge
Antigen PEO1 antibody - middle region
Lot Number QC6322
Method validated Immunocytochemistry
Positive Control Mouse-anti-ds DNA antibody (clone 35I9)
Notes Passed, regarding sensitivity and specificity. The antibody shows a staining pattern consistent with the expected mitochondrial localization of PEO1.
Primary Antibody ABIN2775251
Secondary Antibody Goat-anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 488 conjugate (Invitrogen, A-11034, Lot 702323)
Protocol
  • HeLa cells were grown in DMEM high glucose + 10% FBS on acid-cleaned glass coverslips to 50% confluency.
  • Cells were fixed in 4% para-formaldehyde in growth medium for 25min at RT, then washed once with growth medium and twice with PBS.
  • Permeabilization was achieved by incubation with 0.5% Triton X-100, 10% fetal bovine serum in PBS for 15min.
  • Primary antibodies:
    • ABIN2775251 diluted 1:200 in PBS with 0.1% Triton X-100 , 10% fetal bovine serum.
    • Co-staining of mitochondrial DNA with monoclonal antibody mouse-anti-dsDNA clone 35I9 DNA (Abcam ab27156, Lot GR27035-10) diluted 1:400 in PBS with 0.1% Triton X-100, 10% fetal bovine serum.
  • Incubation for 1h at RT.
  • Wash 3x 5min with PBS.
  • Secondary antibodies:
    • Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor 488 conjugate (Invitrogen, A-11034, Lot 702323) diluted 1:1000 in PBS with 0,1% Triton X-100 , 10% fetal bovine serum, 1µM DAPI.
    • Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor 594 conjugate (Invitrogen, A-11032, Lot 621333).
  • Incubation for 1h at RT.
  • Wash 3x 5min with PBS.
  • The coverslips were mounted with Immu-mount mounting medium (Thermo Fisher, 9990402) and imaged with a 630 x magnification on a Zeiss Axioplan 2 fluorescent microscope using an Axiocam camera and Axiovision 4.8 software.
  • For visualization of PEO1 the excitation wavelength was 488nm and emission wavelength >535nm, exposure time 53ms. DNA was visualized by excitation at 580nm and emission at >590nm, exposure time 410ms.
Validation Images
Immunocytochemistry validation image for anti-Chromosome 10 Open Reading Frame 2 (C10ORF2) (Middle Region) antibody (ABIN2775251) ABIN2775251 was used at a 1:200 dilution to visualize PEO1 (green, secondary antibody...
Immunogen The immunogen is a synthetic peptide directed towards the middle region of human PEO1
Blocking Peptide Blocking peptide for this product available: ABIN981585
Sequence GVFRKFATDN NCHVTLVIHP RKEDDDKELQ TASIFGSAKA SQEADNVLIL
Predicted Reactivity Cow: 100%, Dog: 100%, Guinea Pig: 100%, Horse: 100%, Human: 100%, Mouse: 100%, Rabbit: 100%, Rat: 100%, Zebrafish: 93%
Characteristics This is a rabbit polyclonal antibody against PEO1. It was validated on Western Blot using a cell lysate as a positive control.
Purification Affinity Purified
Antigen
Alternative Name PEO1 (C10ORF2 Antibody Abstract)
Background Twinkle is a mitochondrial protein with structural similarity to the phage T7 primase/helicase (GP4) and other hexameric ring helicases. The twinkle protein colocalizes with mtDNA in mitochondrial nucleoids, and its name derives from the unusual localization pattern reminiscent of twinkling stars.Twinkle is a mitochondrial protein with structural similarity to the phage T7 primase/helicase (GP4) and other hexameric ring helicases. The twinkle protein colocalizes with mtDNA in mitochondrial nucleoids, and its name derives from the unusual localization pattern reminiscent of twinkling stars (Spelbrink et al., 2001 [PubMed 11431692]).[supplied by OMIM]. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications. PRIMARYREFSEQ_SPAN PRIMARY_IDENTIFIER PRIMARY_SPAN COMP 1-213 BG473173.1 6-218 214-1706 BX640829.1 215-1707 1707-3630 BC033762.1 719-2642
Alias Symbols: C10orf2, FLJ21832, PEO, PEOA3, SANDO, TWINL, PEO1, SCA8, ATXN8, IOSCA, MTDPS7
Protein Interaction Partner: ZBTB1, BMI1, SMAD9, ICT1, SQSTM1, AKTIP,
Protein Size: 684
Molecular Weight 77 kDa
Gene ID 56652
NCBI Accession NM_021830, NP_068602
UniProt Q96RR1
Application Notes Optimal working dilutions should be determined experimentally by the investigator.
Comment

Antigen size: 684 AA

Restrictions For Research Use only
Format Liquid
Concentration Lot specific
Buffer Liquid. Purified antibody supplied in 1x PBS buffer with 0.09 % (w/v) sodium azide and 2 % sucrose.
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Avoid repeated freeze-thaw cycles.
Storage -20 °C
Storage Comment For short term use, store at 2-8°C up to 1 week. For long term storage, store at -20°C in small aliquots to prevent freeze-thaw cycles.
Supplier Images
Image no. 1 for anti-Chromosome 10 Open Reading Frame 2 (C10ORF2) (Middle Region) antibody (ABIN2775251) WB Suggested Anti-PEO1 Antibody Titration: 0.2-1 ug/ml ELISA Titer: 1:312500 Posi...
Product cited in: Wang, Xu, Alontaga, Chen, Liu: "Impaired p32 regulation caused by the lymphoma-prone RECQ4 mutation drives mitochondrial dysfunction." in: Cell reports, Vol. 7, Issue 3, pp. 848-58, 2014 (PubMed).

Ylikallio, Tyynismaa, Tsutsui, Ide, Suomalainen: "High mitochondrial DNA copy number has detrimental effects in mice." in: Human molecular genetics, Vol. 19, Issue 13, pp. 2695-705, 2010 (PubMed).

Background publications Farge, Holmlund, Khvorostova, Rofougaran, Hofer, Falkenberg: "The N-terminal domain of TWINKLE contributes to single-stranded DNA binding and DNA helicase activities." in: Nucleic acids research, Vol. 36, Issue 2, pp. 393-403, 2008 (PubMed).

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