Anti-TREX1 antibody [EPR25101-12]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-TREX1 antibody [EPR25101-12] (AB300445)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) cells labelling TREX1 with ab300445 at 1/50 (10.36 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 increased cytoplasmic and nuclear staining in RAW 264.7 cells treated with lipopolysaccharides (LPS, 500 ng/mL) for 24 h is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/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-TREX1 antibody [EPR25101-12] (AB300445)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 500ng/ml LPS for 24 hours (Red) / Untreated control (Green) cells labelling TREX1 with ab300445 at 1/500 dilution (0.1ug) (Red) and Green (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.

• IP

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Immunoprecipitation - Anti-TREX1 antibody [EPR25101-12] (AB300445)

TREX1 was immunoprecipitated from 0.35 mg RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 500ng/ml LPS for 24h whole cell lysate 10 ug with ab300445 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300445 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 500ng/ml LPS for 24h whole cell lysate 10 ug

Lane 2 : ab300445 IP in RAW 264.7 treated with 500ng/ml LPS for 24h whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab300445 in RAW 264.7 treated with 500ng/ml LPS for 24h whole cell lysate

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

Exposure time : 5.5 seconds

All lanes:

Immunoprecipitation - Anti-TREX1 antibody [EPR25101-12] (ab300445) at 1/1000 dilution

All lanes:

RAW 264.7 treated with 500ng/ml LPS for 24h whole cell lysate at 10 µg

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

false

Exposure time: 5.5s

• WB

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Western blot - Anti-TREX1 antibody [EPR25101-12] (AB300445)

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

All lanes:

Western blot - Anti-TREX1 antibody [EPR25101-12] (ab300445) at 1/1000 dilution

Lane 1:

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

Lane 2:

J774A.1 treated with /ml LPS for 18h whole cell lysate 20

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 1/100000 dilution

Observed band size: 30 kDa,33 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-TREX1 antibody [EPR25101-12] (AB300445)

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

All lanes:

Western blot - Anti-TREX1 antibody [EPR25101-12] (ab300445) at 1/1000 dilution

Lane 1:

Untreated RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

Lane 2:

RAW 264.7 treated with /ml LPS for 24h whole cell lysate 20

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 1/100000 dilution

Observed band size: 30 kDa,33 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-TREX1 antibody [EPR25101-12] (AB300445)

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

All lanes:

Western blot - Anti-TREX1 antibody [EPR25101-12] (ab300445) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate

Lane 2:

Mouse thymus tissue lysate

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 1/100000 dilution

Observed band size: 30 kDa,33 kDa

false

Exposure time: 59s