Anti-MBNL1 antibody [EPR26983-11] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MBNL1 antibody [EPR26983-11] - BSA and Azide free (AB309349)

This data was developed using ab309348, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 80% Methanol-fixed, 0.1% TritonX-100 permeabilized Jurkat (human T cell leukemia T lymphocyte) cells labelling MBNL1 with ab309348 at 1/500 (0.976 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing nuclear and weak cytoplasmic staining in Jurkat cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). 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 2ug/ml dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-MBNL1 antibody [EPR26983-11] - BSA and Azide free (AB309349)

This data was developed using ab309348, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Jurkat (human T cell leukemia T lymphocyte from peripheral blood) cells labelling MBNL1 with ab309348 at 1/500 dilution (0.1 ug) (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 1/5000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-MBNL1 antibody [EPR26983-11] - BSA and Azide free (AB309349)

This data was developed using ab309348, the same antibody clone in a different buffer formulation. MBNL1 was immunoprecipitated from 0.35 mg Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate with ab309348 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab309348 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate 10 ug Lane 2 : Jurkat (human t cell leukemia t lymphocyte from peripheral blood) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab309348 in Jurkat whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 3 seconds

All lanes:

Immunoprecipitation - Anti-MBNL1 antibody [EPR26983-11] (<a href='/en-us/products/primary-antibodies/mbnl1-antibody-epr26983-11-ab309348'>ab309348</a>) at 1/30 dilution

All lanes:

Jurkat (human t cell leukemia t lymphocyte from peripheral blood) 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

false

Exposure time: 3s

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-MBNL1 antibody [EPR26983-11] - BSA and Azide free (AB309349)

This data was developed using ab309348, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C2C12 (mouse myoblast) cells labelling MBNL1 with ab309348 at 1/500 dilution (0.1 ug) (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 1/5000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MBNL1 antibody [EPR26983-11] - BSA and Azide free (AB309349)

This data was developed using ab309348, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized C2C12 (mouse myoblasts) cells labelling MBNL1 with ab309348 at 1/500 (0.976 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing nuclear and weak cytoplasmic staining in C2C12 cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). 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 2ug/ml dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-MBNL1 antibody [EPR26983-11] - BSA and Azide free (AB309349)

This data was developed using ab309348, the same antibody clone in a different buffer formulation. MBNL1 was immunoprecipitated from 0.35 mg C2C12 (mouse myoblast) whole cell lysate with ab309348 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab309348 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : C2C12 (mouse myoblast) whole cell lysate 10 ug Lane 2 : C2C12 (mouse myoblast) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab309348 in C2C12 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 15 seconds.

All lanes:

Immunoprecipitation - Anti-MBNL1 antibody [EPR26983-11] (<a href='/en-us/products/primary-antibodies/mbnl1-antibody-epr26983-11-ab309348'>ab309348</a>) at 1/30 dilution

All lanes:

C2C12 (mouse myoblast) 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

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-MBNL1 antibody [EPR26983-11] - BSA and Azide free (AB309349)

This data was developed using ab309348, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : Lanes 1-3 : 6 seconds; Lane 4 : 37 seconds.

All lanes:

Western blot - Anti-MBNL1 antibody [EPR26983-11] (<a href='/en-us/products/primary-antibodies/mbnl1-antibody-epr26983-11-ab309348'>ab309348</a>) at 1/1000 dilution

Lane 1:

Jurkat (human t cell leukemia t lymphocyte from peripheral blood) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

Jurkat transfected with siRNA 1 specifically targeti MBNL1 whole cell lysate at 20 µg

Lane 3:

Jurkat transfected with siRNA 2 specifically targeti MBNL1 whole cell lysate at 20 µg

Lane 4:

HeLa (human cervical adenocarcinoma 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 1/20000 dilution

Observed band size: 42 kDa

false

• WB

Supplier Data

Western blot - Anti-MBNL1 antibody [EPR26983-11] - BSA and Azide free (AB309349)

This data was developed using ab309348, the same antibody clone in a different buffer formulation.

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 23949219 PMID : 16864772).

Exposure time : Lane 1 : 180 seconds; Lane 2 : 136 seconds.

All lanes:

Western blot - Anti-MBNL1 antibody [EPR26983-11] (<a href='/en-us/products/primary-antibodies/mbnl1-antibody-epr26983-11-ab309348'>ab309348</a>) at 1/1000 dilution

Lane 1:

Mouse heart tissue lysate at 20 µg

Lane 2:

C2C12 (mouse myoblast) 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 1/20000 dilution

Observed band size: 42 kDa,38 kDa,34 kDa,26 kDa

false

• WB

Supplier Data

Western blot - Anti-MBNL1 antibody [EPR26983-11] - BSA and Azide free (AB309349)

This data was developed using ab309348, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST This antibody does not cross-react with human MBNL2 and MBNL3. In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 8 seconds.

All lanes:

Western blot - Anti-MBNL1 antibody [EPR26983-11] (<a href='/en-us/products/primary-antibodies/mbnl1-antibody-epr26983-11-ab309348'>ab309348</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag® whole cell lysate at 10 µg

Lane 2:

293T transfected with MBNL1(WT) expression vector containing a myc-His-tag® whole cell lysate at 10 µg

Lane 3:

293T transfected with MBNL2(WT) expression vector containing a myc-His-tag® whole cell lysate at 10 µg

Lane 4:

293T transfected with MBNL3(WT) expression vector containing a myc-His-tag® whole cell lysate 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 1/100000 dilution

Observed band size: 42 kDa,38 kDa,34 kDa,26 kDa

false

Exposure time: 8s