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  1. Link

    tms1asc-antibody-epr23978-28-ab283684.pdf

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Cell Biology Apoptosis Intracellular Associated Proteins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Western blot - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
  • Western blot - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
  • Immunocytochemistry/ Immunofluorescence - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
  • Flow Cytometry - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
  • Immunoprecipitation - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
  • Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23978-28] to TMS1/ASC
  • Suitable for: ICC/IF, WB, Flow Cyt, IP, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free

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Protein
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Recombinant Human TMS1/ASC protein (ab131786)
Primary
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Anti-TMS1/ASC antibody [EPR23978-28] - BSA and Azide free (ab283712)
Lysate
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TMS1 overexpression 293T lysate (whole cell) (ab94129)

View more associated products

Overview

  • Product name

    Anti-TMS1/ASC antibody [EPR23978-28]
    See all TMS1/ASC primary antibodies
  • Description

    Rabbit monoclonal [EPR23978-28] to TMS1/ASC
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, Flow Cyt, IP, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human spleen, colon and tonsil tissue lysate; Wild-type HAP1 and THP-1 cell lysates. IHC-P: Human tonsil, colon and colon cancer tissue. ICC/IF: THP-1 cells. Flow Cyt: HeLa cells. IP: THP-1 cells.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR23978-28
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Associated Proteins
    • Cell Biology
    • Apoptosis
    • Intracellular
    • Caspases etc
    • CARD Family
    • Cancer
    • Tumor biomarkers
    • Other
    • Neuroscience
    • Processes

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab283684 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
ICC/IF
1/100.
WB
1/1000. Predicted molecular weight: 22 kDa.
Flow Cyt
1/5000.
IP
1/30.
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
ICC/IF
1/100.
WB
1/1000. Predicted molecular weight: 22 kDa.
Flow Cyt
1/5000.
IP
1/30.
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Promotes caspase-mediated apoptosis. This proapoptotic activity is mediated predominantly through the activation of caspase-9. May be a component of the inflammasome, a protein complex which also includes NALP2, CARD8 and CASP1 and whose function would be the activation of proinflammatory caspases.
  • Tissue specificity

    Widely expressed at low levels. Detected in peripheral blood leukocytes, lung, small intestine, spleen, thymus, colon and at lower levels in placenta, liver and kidney. Very low expression in skeletal muscle, heart and brain. Detected in the leukemia cell lines HL-60 and U937, but not in Jurkat T-cell lymphoma and Daudi Burkitt's lymphoma. Detected in the melanoma cell line WM35, but not in WM793. Not detected in HeLa cervical carcinoma cells and Molt 4 lymphocytic leukemia cells.
  • Sequence similarities

    Contains 1 CARD domain.
    Contains 1 DAPIN domain.
  • Domain

    Interacts with CIAS1/PYPAF1 and PYDC1 via the DAPIN domain.
  • Post-translational
    modifications

    Phosphorylated.
  • Cellular localization

    Cytoplasm. Upstream of caspase activation, a redistribution from the cytoplasm to the aggregates occurs. These appear as hollow, perinuclear spherical, ball-like structures.
  • Target information above from: UniProt accession Q9ULZ3 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 29108 Human
    • Omim: 606838 Human
    • SwissProt: Q9ULZ3 Human
    • Unigene: 499094 Human
    • Alternative names

      • Apoptosis associated speck like protein containing a CARD antibody
      • Apoptosis-associated speck-like protein containing a CARD antibody
      • ASC antibody
      • ASC_HUMAN antibody
      • CARD 5 antibody
      • CARD5 antibody
      • Caspase recruitment domain containing protein 5 antibody
      • Caspase recruitment domain protein 5 antibody
      • Caspase recruitment domain-containing protein 5 antibody
      • hASC antibody
      • MGC10332 antibody
      • PYCARD antibody
      • PYD and CARD domain containing antibody
      • PYD and CARD domain containing protein antibody
      • PYD and CARD domain-containing protein antibody
      • Target of methylation induced silencing 1 antibody
      • Target of methylation-induced silencing 1 antibody
      • TMS 1 antibody
      • TMS antibody
      • TMS1 antibody
      see all

    Images

    • Western blot - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
      Western blot - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
      All lanes : Anti-TMS1/ASC antibody [EPR23978-28] (ab283684) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 cell lysate
      Lane 2 : TMS1/ASC knockout HAP1 cell lysate
      Lane 3 : THP-1 (human monocytic leukemia monocyte) whole cell lysate
      Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution

      Predicted band size: 22 kDa
      Observed band size: 17,24 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer and concentration: 3% NFDM/TBST

      Negative control: HeLa (PMID:11103776;10567338).

      Lanes 1-4: Merged signal (red and green). Green - ab283684 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa.

      Ab283684 Anti-TMS1/ASC antibody [EPR23978-28] was shown to specifically react with TMS1/ASC in wild-type HAP1 cells. Loss of signal was observed when knockout cell line (knockout cell lysate) was used. Wild-type and TMS1/ASC knockout samples were subjected to SDS-PAGE. ab283684 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging. 

       

    • Western blot - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
      Western blot - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
      All lanes : Anti-TMS1/ASC antibody [EPR23978-28] (ab283684) at 1/1000 dilution

      Lane 1 : Human spleen tissue lysate
      Lane 2 : Human colon tissue lysate
      Lane 3 : Human tonsil tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Predicted band size: 22 kDa
      Observed band size: 12,17,24 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer and concentration: 5% NFDM/TBST

      ASC has four isoforms. 24-kDa ASC, 17-kDa ASC-b and 12-kDa ASC-C are observed. The molecular weight observed is consistent with what has been described in the literature (PMID:20482797).

      Exposure time: 81 seconds

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)

      Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling TMS1/ASC with ab283684 at 1/2000 (0.247 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human tonsil. The section was incubated with ab283684 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)

      Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling TMS1/ASC with ab283684 at 1/2000 (0.247 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human colon. The section was incubated with ab283684 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)

      Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labelling TMS1/ASC with ab283684 at 1/2000 (0.247 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human colon cancer (PMID: 14643031). The section was incubated with ab283684 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

    • Immunocytochemistry/ Immunofluorescence - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
      Immunocytochemistry/ Immunofluorescence - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized THP-1 cells labelling TMS1/ASC with ab283684 at 1/100 (4.94 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic taining in THP-1 cell line. Negative control: HeLa (PMID: 11103776). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

    • Flow Cytometry - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
      Flow Cytometry - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)

      Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell, Left) / THP-1 (Human monocytic leukemia monocyte, Right) cells labelling TMS1/ASC with ab283684 at 1/5000 dilution (0.01ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control: HeLa (PMID: 11103776).

    • Immunoprecipitation - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
      Immunoprecipitation - Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)

      TMS1/ASC was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte) whole cell lysate 20 ug with ab283684 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283684 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: THP-1 (human monocytic leukemia monocyte) whole cell lysate 20 ug

      Lane 2: ab283684 IP in THP-1 whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab283684 in THP-1 whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 10 seconds

       

    • Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)
      Anti-TMS1/ASC antibody [EPR23978-28] (ab283684)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (0)

    Publishing research using ab283684? Please let us know so that we can cite the reference in this datasheet.

    ab283684 has not yet been referenced specifically in any publications.

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