Recombinant Anti-TMS1/ASC antibody [EPR23978-28] - BSA and Azide free (ab283712)
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 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-TMS1/ASC antibody [EPR23978-28] - BSA and Azide free
See all TMS1/ASC primary antibodies -
Description
Rabbit monoclonal [EPR23978-28] to TMS1/ASC - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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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.
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General notes
ab283712 is the carrier-free version of ab283684.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. -
Storage buffer
pH: 7.2
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23978-28 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab283712 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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 22 kDa.
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 22 kDa. |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
Target
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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
modificationsPhosphorylated. -
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. - Information by UniProt
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Database links
- Entrez Gene: 29108 Human
- Omim: 606838 Human
- SwissProt: Q9ULZ3 Human
- Unigene: 499094 Human
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Alternative names
- Apoptosis associated speck like protein containing a CARD antibody
- Apoptosis-associated speck-like protein containing a CARD antibody
- ASC antibody
see all
Images
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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 kDaThis data was developed using ab283684, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 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.
Exposure time:
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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?This data was developed using ab283684, the same antibody clone in a different buffer formulation.
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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR23978-28] - BSA and Azide free (ab283712)
This data was developed using ab283684, the same antibody clone in a different buffer formulation.
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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR23978-28] - BSA and Azide free (ab283712)
This data was developed using ab283684, the same antibody clone in a different buffer formulation.
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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR23978-28] - BSA and Azide free (ab283712)
This data was developed using ab283684, the same antibody clone in a different buffer formulation.
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
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Immunocytochemistry/ Immunofluorescence - Anti-TMS1/ASC antibody [EPR23978-28] - BSA and Azide free (ab283712)
This data was developed using ab283684, the same antibody clone in a different buffer formulation.
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.
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Flow Cytometry (Intracellular) - Anti-TMS1/ASC antibody [EPR23978-28] - BSA and Azide free (ab283712)
This data was developed using ab283684, the same antibody clone in a different buffer formulation.
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).
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This data was developed using ab283684, the same antibody clone in a different buffer formulation.
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
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.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab283712 has not yet been referenced specifically in any publications.