Anti-Pyrin antibody [EPR18675]

• IP

Lab

Immunoprecipitation - Anti-Pyrin antibody [EPR18675] (AB214772)

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

VeriBlot for IP secondary antibody(HRP)(ab131366) ws used as a secondary.

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

All lanes:

Immunoprecipitation - Anti-Pyrin antibody [EPR18675] (ab214772) at 1/1000 dilution

Lane 1:

Mouse spleen tissue whole cell lysate at 10 µg

Lane 2:

ab214772 at 1/30 IP in Mouse spleen tissue whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab214772 in mouse spleen tissue 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

Observed band size: 86-110 kDa

true

Exposure time: 41s

• WB

Supplier Data

Western blot - Anti-Pyrin antibody [EPR18675] (AB214772)

Blocking/Dilution buffer : 5% NFDM/TBST.

The lysates were kindly provided by our collaborator Dr Feng Shao's lab, NIBS.

All lanes:

Western blot - Anti-Pyrin antibody [EPR18675] (ab214772) at 1/1000 dilution

Lane 1:

Bone marrow derived-macrophage of Pyrin -/- mice at 10 µg

Lane 2:

Bone marrow derived-macrophage of Pyrin -/- mice stimulated with LPS at 10 µg

Lane 3:

Bone marrow derived-macrophage of wild type C57/B6 mice at 10 µg

Lane 4:

Bone marrow derived-macrophage of wild type C57/B6 mice stimulated with LPS at 10 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

Predicted band size: 86 kDa

Observed band size: 110 kDa

false

Exposure time: 1min

• WB

Lab

Western blot - Anti-Pyrin antibody [EPR18675] (AB214772)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Negative control : heart.

To minimize protein degradation, cells/tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The identity of the band between 75 kDa and 50 kDa is unknown.

All lanes:

Western blot - Anti-Pyrin antibody [EPR18675] (ab214772) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 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: 86-110 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Pyrin antibody [EPR18675] (AB214772)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Negative control : lung, skeletal muscle and kidney.

To minimize protein degradation, cells/tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

All lanes:

Western blot - Anti-Pyrin antibody [EPR18675] (ab214772) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse lung tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 4:

Mouse kidney tissue lysate at 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: 86-110 kDa

false

Exposure time: 180s