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  1. Link

    tlsfus-antibody-cl0190-ab154141.pdf

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Epigenetics and Nuclear Signaling Transcription Cancer susceptibility Proto-oncogenes
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Validated using a knockout cell line

Anti-TLS/FUS antibody [CL0190] (ab154141)

  • Datasheet
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Western blot - Anti-TLS/FUS antibody [CL0190] (ab154141)
  • Western blot - Anti-TLS/FUS antibody [CL0190] (ab154141)
  • Immunocytochemistry/ Immunofluorescence - Anti-TLS/FUS antibody [CL0190] (ab154141)
  • Western blot - Anti-TLS/FUS antibody [CL0190] (ab154141)
  • Immunocytochemistry/ Immunofluorescence - Anti-TLS/FUS antibody [CL0190] (ab154141)
  • Western blot - Anti-TLS/FUS antibody [CL0190] (ab154141)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [CL0190] (ab154141)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [CL0190] (ab154141)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [CL0190] (ab154141)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [CL0190] (ab154141)

Key features and details

  • Mouse monoclonal [CL0190] to TLS/FUS
  • Suitable for: IHC-P, WB, ICC/IF
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

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Overview

  • Product name

    Anti-TLS/FUS antibody [CL0190]
    See all TLS/FUS primary antibodies
  • Description

    Mouse monoclonal [CL0190] to TLS/FUS
  • Host species

    Mouse
  • Tested applications

    Suitable for: IHC-P, WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Recombinant fragment, corresponding to amino acids 86-213 of Human TLS/FUS.

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control

    • IHC-P: Human kidney, stomach, prostate and pancreas tissues. WB: Hap1, HEK-293T, K562, HepG2 and U-251 cell lysates. ICC/IF: HeLa cells.
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.02% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine)
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    CL0190
  • Isotype

    IgG1
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Cancer susceptibility
    • Proto-oncogenes
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Cancer
    • Signal transduction
    • Other
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Transcription factors
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Other

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
  • KO cell lines

    • Human FUS (TLS) knockout HEK-293T cell line (ab266587)
  • KO cell lysates

    • Human FUS (TLS) knockout HEK-293T cell lysate (ab257100)
  • KO cell pellets

    • Human FUS (TLS) knockout HEK-293T cell pellet (ab278884)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab154141 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P
1/2500 - 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
1/500 - 1/1000. Predicted molecular weight: 53 kDa.
ICC/IF
Use a concentration of 1 - 4 µg/ml.

Fixation/Permeabilization: PFA/Triton X-100.

Notes
IHC-P
1/2500 - 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
1/500 - 1/1000. Predicted molecular weight: 53 kDa.
ICC/IF
Use a concentration of 1 - 4 µg/ml.

Fixation/Permeabilization: PFA/Triton X-100.

Target

  • Function

    Binds both single-stranded and double-stranded DNA and promotes ATP-independent annealing of complementary single-stranded DNAs and D-loop formation in superhelical double-stranded DNA. May play a role in maintenance of genomic integrity.
  • Tissue specificity

    Ubiquitous.
  • Involvement in disease

    Note=A chromosomal aberration involving FUS is found in a patient with malignant myxoid liposarcoma. Translocation t(12;16)(q13;p11) with DDIT3.
    Note=A chromosomal aberration involving FUS is a cause of acute myeloid leukemia (AML). Translocation t(16;21)(p11;q22) with ERG.
    Defects in FUS may be a cause of angiomatoid fibrous histiocytoma (AFH) [MIM:612160]. A distinct variant of malignant fibrous histiocytoma that typically occurs in children and adolescents and is manifest by nodular subcutaneous growth. Characteristic microscopic features include lobulated sheets of histiocyte-like cells intimately associated with areas of hemorrhage and cystic pseudovascular spaces, as well as a striking cuffing of inflammatory cells, mimicking a lymph node metastasis. Note=A chromosomal aberration involving FUS is found in a patient with angiomatoid fibrous histiocytoma. Translocation t(12;16)(q13;p11.2) with ATF1 generates a chimeric FUS/ATF1 protein.
    Defects in FUS are the cause of amyotrophic lateral sclerosis type 6 (ALS6) [MIM:608030]. ALS6 is a familial form of amyotrophic lateral sclerosis. ALS is a neurodegenerative disorder affecting upper motor neurons in the brain and lower motor neurons in the brain stem and spinal cord, resulting in fatal paralysis. Sensory abnormalities are absent. Death usually occurs within 2 to 5 years. The etiology of amyotrophic lateral sclerosis is likely to be multifactorial, involving both genetic and environmental factors. The disease is inherited in 5-10%.
  • Sequence similarities

    Belongs to the RRM TET family.
    Contains 1 RanBP2-type zinc finger.
    Contains 1 RRM (RNA recognition motif) domain.
  • Post-translational
    modifications

    Arg-216 and Arg-218 are dimethylated, probably to asymmetric dimethylarginine.
  • Cellular localization

    Nucleus.
  • Target information above from: UniProt accession P35637 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2521 Human
    • Entrez Gene: 233908 Mouse
    • Entrez Gene: 317385 Rat
    • Omim: 137070 Human
    • SwissProt: P35637 Human
    • SwissProt: P56959 Mouse
    • Unigene: 46894 Human
    • Unigene: 277680 Mouse
    • Alternative names

      • 75 kDa DNA pairing protein antibody
      • 75 kDa DNA-pairing protein antibody
      • ALS6 antibody
      • Amyotrophic lateral sclerosis 6 antibody
      • fus antibody
      • FUS CHOP antibody
      • Fus like protein antibody
      • FUS_HUMAN antibody
      • FUS1 antibody
      • Fused in sarcoma antibody
      • Fusion (involved in t(12;16) in malignant liposarcoma) antibody
      • Fusion derived from t(12;16) malignant liposarcoma antibody
      • Fusion gene in myxoid liposarcoma antibody
      • Heterogeneous nuclear ribonucleoprotein P2 antibody
      • hnRNP P2 antibody
      • hnRNPP2 antibody
      • Oncogene FUS antibody
      • Oncogene TLS antibody
      • POMp75 antibody
      • RNA binding protein FUS antibody
      • RNA-binding protein FUS antibody
      • TLS antibody
      • TLS CHOP antibody
      • Translocated in liposarcoma antibody
      • Translocated in liposarcoma protein antibody
      see all

    Images

    • Western blot - Anti-TLS/FUS antibody [CL0190] (ab154141)
      Western blot - Anti-TLS/FUS antibody [CL0190] (ab154141)

      ab154141 was shown to react with FUS in wild-type HeLa cells in Western blot with loss of signal observed in a FUS knockout cell line. Wild-type HeLa and FUS knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab154141 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 1/5000 before imaging. These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

    • Western blot - Anti-TLS/FUS antibody [CL0190] (ab154141)
      Western blot - Anti-TLS/FUS antibody [CL0190] (ab154141)
      All lanes : Anti-TLS/FUS antibody [CL0190] (ab154141) at 1/500 dilution

      Lane 1 : Wild-type HAP1 cell lysate
      Lane 2 : TLS/FUS knockout HAP1 cell lysate
      Lane 3 : K562 cell lysate
      Lane 4 : HepG2 cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 53 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab154141 observed at 80 kDa. Red - loading control, ab181602, observed at 37 kDa.

      ab154141 was shown to specifically react with TLS/FUS when TLS/FUS knockout samples were used. Wild-type and TLS/FUS knockout samples were subjected to SDS-PAGE. ab154141 and ab181602 (loading control to GAPDH) were diluted at 1/500 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD)preadsorbed (ab216777) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-TLS/FUS antibody [CL0190] (ab154141)
      Immunocytochemistry/ Immunofluorescence - Anti-TLS/FUS antibody [CL0190] (ab154141)

      Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling TLS/FUS with ab154141 showing nuclear (without nucleoli) staining in green. Microtubule are visualized in red.

    • Western blot - Anti-TLS/FUS antibody [CL0190] (ab154141)
      Western blot - Anti-TLS/FUS antibody [CL0190] (ab154141)
      All lanes : Anti-TLS/FUS antibody [CL0190] (ab154141) at 1/500 dilution

      Lane 1 : Wild-type HEK-293T cell lysate
      Lane 2 : FUS knockout HEK-293T cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 53 kDa
      Observed band size: 80 kDa why is the actual band size different from the predicted?



      Lanes 1- 2: Merged signal (red and green). Green - ab154141 observed at 80 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.

       ab154141 was shown to react with TLS/FUS in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266587 (knockout cell lysate ab257100) was used. Wild-type HEK-293T and FUS knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab154141 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-TLS/FUS antibody [CL0190] (ab154141)
      Immunocytochemistry/ Immunofluorescence - Anti-TLS/FUS antibody [CL0190] (ab154141)
      ab154141 was shown to react with FUS in wild-type HeLa cells in Immunocytochemistry with loss of signal observed in a FUS knockout cell line. Wild-type and Knockout cells were mixed and pelleted at a 1:1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 5%BSA+5%goat serum (30min). The cells were then incubated with ab154141 at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-mouse secondary antibody to (Alexa Fluor® 555) at 0.5 μg/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (knockout) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and knockout cells are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880).

      These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
    • Western blot - Anti-TLS/FUS antibody [CL0190] (ab154141)
      Western blot - Anti-TLS/FUS antibody [CL0190] (ab154141)
      All lanes : Anti-TLS/FUS antibody [CL0190] (ab154141)

      Lane 1 : U-251 cells, transfected with control siRNA
      Lane 2 : U-251 cells, transfected with target specific siRNA probe #1
      Lane 3 : U-251 cells, transfected with target specific siRNA probe #2

      Predicted band size: 53 kDa



      Downregulation of antibody signal confirms target specificity. Remaining % intensity, relative control lane, is indicated. Anti-GAPDH monoclonal antibody was used as loading control.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [CL0190] (ab154141)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [CL0190] (ab154141)
      Immunohistochemical analysis of Human kidney tissue labeling TLS/FUS with ab154141 at 1/2500 dilution.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [CL0190] (ab154141)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [CL0190] (ab154141)
      Immunohistochemical analysis of Human prostate tissue labeling TLS/FUS with ab154141 at 1/2500 dilution.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [CL0190] (ab154141)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [CL0190] (ab154141)
      Immunohistochemical analysis of Human stomach tissue labeling TLS/FUS with ab154141 at 1/2500 dilution.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [CL0190] (ab154141)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TLS/FUS antibody [CL0190] (ab154141)
      Immunohistochemical analysis of Human pancreas tissue labeling TLS/FUS with ab154141 at 1/2500 dilution.

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (2)

    Publishing research using ab154141? Please let us know so that we can cite the reference in this datasheet.

    ab154141 has been referenced in 2 publications.

    • Kumar-Singh A  et al. Mapping the Interactome of the Nuclear Heparan Sulfate Proteoglycan Syndecan-1 in Mesothelioma Cells. Biomolecules 10:N/A (2020). PubMed: 32664515
    • Owen I  et al. The prion-like domain of Fused in Sarcoma is phosphorylated by multiple kinases affecting liquid- and solid-phase transitions. Mol Biol Cell 31:2522-2536 (2020). PubMed: 32877292

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Immunocytochemistry/ Immunofluorescence abreview for Anti-TLS/FUS antibody [CL0190]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (mouse embryonic fibroblasts)
    Permeabilization
    Yes - 0.2% Triton-X
    Specification
    mouse embryonic fibroblasts
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 23°C
    Fixative
    Paraformaldehyde
    Read More

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    Verified customer

    Submitted Dec 08 2020

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