Anti-Filamin B antibody [EPR24525-3]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Filamin B antibody [EPR24525-3] (AB282106)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HT-1080 (Human fibrosarcoma epithelial cell) cells labelling Filamin B with ab282106 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, ab150077) at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Filamin B antibody [EPR24525-3] (AB282106)

Flow cytometric analysis of 4% paraformaldehyde-fixed 90% methanol permeabilized 293T (Human embryonic kidney epithelial cell) cells labelling Filamin B with ab282106 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, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Filamin B antibody [EPR24525-3] (AB282106)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling Filamin B with ab282106 at 1/5000 (0.103 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human colon. (PMID : 32907537) The section was incubated with ab282106 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Filamin B antibody [EPR24525-3] (AB282106)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HT-1080 cells labelling Filamin B with ab282106 at 1/50 (10.3 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluo^r® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoskeletal staining in HT-1080 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Filamin B antibody [EPR24525-3] (AB282106)

Immunohistochemical analysis of paraffin-embedded Human skin tissue labelling Filamin B with ab282106 at 1/5000 (0.103 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human skin. The section was incubated with ab282106 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Filamin B antibody [EPR24525-3] (AB282106)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized C2C12 (Mouse myoblasts myoblast) cells labelling Filamin B with ab282106 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, ab150077) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Filamin B antibody [EPR24525-3] (AB282106)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C2C12 cells labelling Filamin B with ab282106 at 1/50 (10.3 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoskeletal staining in C2C12 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Filamin B antibody [EPR24525-3] (AB282106)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labelling Filamin B with ab282106 at 1/5000 (0.103 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat colon. The section was incubated with ab282106 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Filamin B antibody [EPR24525-3] (AB282106)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labelling Filamin B with ab282106 at 1/5000 (0.103 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on Mouse colon. The section was incubated with ab282106 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• WB

Lab

Western blot - Anti-Filamin B antibody [EPR24525-3] (AB282106)

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

The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID : 28175289, 11807098).

Low expression : skeletal muscle (PMID : 28175289, 11807098).

Exposure time : 26 seconds.

All lanes:

Western blot - Anti-Filamin B antibody [EPR24525-3] (ab282106) at 1/1000 dilution

Lane 1:

Human lung tissue lysate at 20 µg

Lane 2:

Human heart tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Lane 4:

Mouse lung tissue lysate at 20 µg

Lane 5:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 6:

Mouse brain 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 1/50000 dilution

Predicted band size: 278 kDa

Observed band size: 315 kDa

false

• WB

Lab

Western blot - Anti-Filamin B antibody [EPR24525-3] (AB282106)

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

The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID : 28175289, 11807098).

Lysates were made freshly and used in WB test immediately to minimize protein degradation.

Exposure time : Lanes 1-2 : 15 seconds; lanes 3-4 : 8 seconds.

All lanes:

Western blot - Anti-Filamin B antibody [EPR24525-3] (ab282106) at 1/1000 dilution

Lane 1:

A-204 (human muscle rhabdomyosarcoma) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

A375 (human malignant melanoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HT-1080 (human fibrosarcoma epithelial cell) whole cell 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 1/50000 dilution

Predicted band size: 278 kDa

Observed band size: 315 kDa

false

• WB

Lab

Western blot - Anti-Filamin B antibody [EPR24525-3] (AB282106)

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

The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID : 28175289, 11807098).

Exposure time : 3 seconds.

All lanes:

Western blot - Anti-Filamin B antibody [EPR24525-3] (ab282106) at 1/1000 dilution

Lane 1:

Mouse colon tissue lysate at 20 µg

Lane 2:

Mouse stomach 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 1/50000 dilution

Predicted band size: 278 kDa

Observed band size: 315 kDa

false

• WB

Lab

Western blot - Anti-Filamin B antibody [EPR24525-3] (AB282106)

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

The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID : 28175289, 11807098).

Low expression : skeletal muscle (PMID : 28175289, 11807098).

Exposure time : 10 seconds.

All lanes:

Western blot - Anti-Filamin B antibody [EPR24525-3] (ab282106) at 1/1000 dilution

Lane 1:

Rat lung tissue lysate at 20 µg

Lane 2:

Rat skeletal muscle 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 1/50000 dilution

Predicted band size: 278 kDa

Observed band size: 315 kDa

false