Anti-TMS1/ASC antibody [EPR28201-61]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-TMS1/ASC antibody [EPR28201-61] (AB307560)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeABilized J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) cells lABelling TMS1/ASC with ab307560 at 1/500 (0.968 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 nuclear and cytoplasmic staining in J774A.1 cell line. Negative control : RAW 264.7 (PMID : 28766178Ј©Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. 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 [EPR28201-61] (AB307560)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW264.7 (mouse abelson murine leukemia virus-induced tumor macrophage, Left) / J774A.1 (mouse reticulum cell sarcoma monocyte macrophage, Right) cells labelling TMS1/ASC with ab307560 at 1/500 dilution (0.1ug) (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 : RAW 264.7 (PMID : 28766178).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR28201-61] (AB307560)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue lABeling TMS1/ASC with ab307560 at 1/1000 (0.484 ug/ml) followed by a ready to use LeicaDS9800 (Bond? Polymer Refine Detection) was used. Low expression tissue : no staining on rat skeletal muscle.The section was incubated with ab307560 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR28201-61] (AB307560)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue lABeling TMS1/ASC with ab307560 at 1/1000 (0.484 ug/ml) followed by a ready to use LeicaDS9800 (Bond? Polymer Refine Detection) was used. Positive staining on mouse spleen.The section was incubated with ab307560 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR28201-61] (AB307560)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue lABeling TMS1/ASC with ab307560 at 1/1000 (0.484 ug/ml) followed by a ready to use LeicaDS9800 (Bond? Polymer Refine Detection) was used. Positive staining on mouse colon.The section was incubated with ab307560 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR28201-61] (AB307560)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue lABeling TMS1/ASC with ab307560 at 1/1000 (0.484 ug/ml) followed by a ready to use LeicaDS9800 (Bond? Polymer Refine Detection) was used. Positive staining on rat spleen.The section was incubated with ab307560 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR28201-61] (AB307560)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue lABeling TMS1/ASC with ab307560 at 1/1000 (0.484 ug/ml) followed by a ready to use LeicaDS9800 (Bond? Polymer Refine Detection) was used. Positive staining on rat colon.The section was incubated with ab307560 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR28201-61] (AB307560)

Immunohistochemical analysis of paraffin-embedded Mouse head and neck tissue lABeling TMS1/ASC with ab307560 at 1/1000 (0.484 ug/ml) followed by a ready to use LeicaDS9800 (Bond? Polymer Refine Detection) was used. Positive staining on mouse head and neck squamous cell carcinoma.The section was incubated with ab307560 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR28201-61] (AB307560)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscl tissue lABeling TMS1/ASC with ab307560 at 1/1000 (0.484 ug/ml) followed by a ready to use LeicaDS9800 (Bond? Polymer Refine Detection) was used. Low expression tissue : no staining on mouse skeletal muscle.The section was incubated with ab307560 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMS1/ASC antibody [EPR28201-61] (AB307560)

Immunohistochemical analysis of paraffin-embedded (A) J774A.1 (mouse r tissue lABeling TMS1/ASC with ab307560 at 1/1000 (0.484 ug/ml) followed by a ready to use LeicaDS9800 (Bond? Polymer Refine Detection) was used. Positive staining on (A) J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) cell pellet, no staining on (B) RAW 264.7 (Mouse ABelson murine leukemia virus-induced tumor macrophage) cell pellet.The section was incubated with ab307560 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

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Immunoprecipitation - Anti-TMS1/ASC antibody [EPR28201-61] (AB307560)

TMS1/ASC was immunoprecipitated from 0.35 mg J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage ) whole cell lysate 10 ug with ab307560 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307560 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage ) whole cell lysate 10 ug
Lane 2 : ab307560 IP in J774A.1 whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307560 in J774A.1 whole cell lysate

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

Exposure time : 50 seconds

All lanes:

Immunoprecipitation - Anti-TMS1/ASC antibody [EPR28201-61] (ab307560) at 1/1000 dilution

Lane 1:

J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage ) whole cell lysate 10 μg

Lane 2:

J774A.1 whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 21 kDa

false

Exposure time: 50s

• WB

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Western blot - Anti-TMS1/ASC antibody [EPR28201-61] (AB307560)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTNegative control : RAW 264.7 (PMID : 28766178); Low expression tissue : skeletal muscle (PMID : 32659370). Exposure time : Lanes 1-6 : 15 seconds; Lane 7 : 125 seconds.

All lanes:

Western blot - Anti-TMS1/ASC antibody [EPR28201-61] (ab307560) at 1/1000 dilution

Lane 1:

J774A.1 (mouse reticum cell sarcoma monocyte/macrophage ) whole cell lysate 20 μg

Lane 2:

RAW 264.7 (mouse elson murine leukemia virus-induced tumor macrophage) whole cell lysate 20 μg

Lane 3:

Mouse spleen tissue lysate 20 μg

Lane 4:

Mouse colon tissue lysate 20 μg

Lane 5:

Mouse skeletal muscle tissue lysate 20 μg

Lane 6:

Rat spleen tissue lysate 20 μg

Lane 7:

Rat colon tissue lysate 20 μg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 21 kDa

false

Exposure time: 15s