Anti-TMS1/ASC antibody [EPR23978-28] - BSA and Azide free

• ICC/IF

Supplier Data

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.

• Flow Cyt (Intra)

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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).

• IHC-P

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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

• IHC-P

Supplier Data

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

• IHC-P

Supplier Data

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

• IP

Supplier Data

Immunoprecipitation - 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.

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

All lanes:

Immunoprecipitation - Anti-TMS1/ASC antibody [EPR23978-28] (<a href='/en-us/products/primary-antibodies/tms1-asc-antibody-epr23978-28-ab283684'>ab283684</a>)

Predicted band size: 22 kDa

Observed band size: 12 kDa,17 kDa,24 kDa

false

• WB

Lab

Western blot - 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.

Western blot : Rabbit Monoclonal [EPR23978-28] to TMS1/ASC ab283684 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 22 kDa in Wild-type A549 cell lysates with no signal observed at this size in PYCARD knockout A549 cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.

Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-TMS1/ASC antibody [EPR23978-28] (<a href='/en-us/products/primary-antibodies/tms1-asc-antibody-epr23978-28-ab283684'>ab283684</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

PYCARD knockout A549 at 20 µg

Lane 3:

HT-29 at 20 µg

Lane 4:

HEK-293 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 22 kDa

Observed band size: 22 kDa

false

• WB

Lab

Western blot - 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.

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

All lanes:

Western blot - Anti-TMS1/ASC antibody [EPR23978-28] (<a href='/en-us/products/primary-antibodies/tms1-asc-antibody-epr23978-28-ab283684'>ab283684</a>) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 20 µg

Lane 2:

Human colon tissue lysate at 20 µg

Lane 3:

Human tonsil tissue lysate at 20 µg

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 kDa,17 kDa,24 kDa

false