Anti-CARD9 antibody [EPR28960-91]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-CARD9 antibody [EPR28960-91] (AB322352)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A549 (human lung carcinoma epithelial cell, Left) / THP-1 (human monocytic leukemia monocyte, Right) cells labelling CARD9 with ab322352 at 1/5000 (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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081 at 1/5000 was used as the secondary antibody.

Low expression : A549.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-CARD9 antibody [EPR28960-91] (AB322352)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MEF (mouse embryo fibroblast, Left)/ RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage, Right) cells labelling CARD9 with ab322352 at 1/5000 (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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081 at 1/5000 was used as the secondary antibody.

Negative Control : MEF.

• WB

Supplier Data

Western blot - Anti-CARD9 antibody [EPR28960-91] (AB322352)

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

Low expression : A549 (PMID : 32164050 ), 293T (PMID : 33558980).

The expression profile/ molecular weight observed is consistent with what has been described in the literature(PMID : 32164050), This antibody can recognize 3 isoforms of CARD9 and isoforms have different molecular weight.

The identity of the higher MW band at 150 kDa (in lane 1) is unknown.

The identity of the lower MW band at 25 kDa (in lane 5) is unknown.

This blot of lane 1 and lane 5 was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

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

All lanes:

Western blot - Anti-CARD9 antibody [EPR28960-91] (ab322352) at 1/1000 dilution

Lane 1:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 2:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 3:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 5:

Human bone marrow tissue lysate at 40 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: 43-62 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CARD9 antibody [EPR28960-91] (AB322352)

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

Negative control : MEF.

Low expression : skeletal muscle (PMID : 11053425).

The expression profile/ molecular weight observed is consistent with what has been described in the literature(PMID : 32164050), This antibody can recognize 3 isoforms of CARD9 and isoforms have different molecular weight.

The identity of the lower MW band at about 30 kDa (in lane 1) is unknown.

The identity of the higher MW bands at about 100 kDa and the lower MW bands at about 40 kDa (in lane 4) are unknown.

This blot of lanes 3-4 was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

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

All lanes:

Western blot - Anti-CARD9 antibody [EPR28960-91] (ab322352) at 1/1000 dilution

Lane 1:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 40 µg

Lane 2:

MEF (mouse embryo fibroblast) whole cell lysate at 40 µg

Lane 3:

Mouse spleen tissue lysate at 40 µg

Lane 4:

Mouse skeletal muscle lysate at 40 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: 43-62 kDa,36 kDa

false

Exposure time: 136s