Anti-non-muscle Myosin IIB/MYH10 antibody [EPR22564-23] - Goat IgG (Chimeric) - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-non-muscle Myosin IIB/MYH10 antibody [EPR22564-23] - Goat IgG (Chimeric) - BSA and Azide free (AB300649)

Immunohistochemical analysis of paraffin-embedded Human cerebellum tissue labeling non-muscle Myosin IIB/MYH10 with ab300649 at 1/500 (1.96 ug/ml) followed by a Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution. Positive staining on human cerebellum. The section was incubated with ab300649 for 30 mins at room temperature, the 2nd antibody Donkey Anti-Goat IgG (HRP) ab205723 was used.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IP

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Immunoprecipitation - Anti-non-muscle Myosin IIB/MYH10 antibody [EPR22564-23] - Goat IgG (Chimeric) - BSA and Azide free (AB300649)

Non-muscle Myosin IIB/MYH10 was immunoprecipitated from 0.35 mg HEK-293T (human embryonic kidney epithelial cell) whole cell lysate, with ab300649 at 1/30 dilution (2 μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300649 at 1/1000 dilution. Peroxidase conjugated Rabbit anti-Goat IgG (H+L), was used as the secondary antibody at 1/5000 dilution.

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

Exposure time : 3.25 second

All lanes:

Immunoprecipitation - Anti-non-muscle Myosin IIB/MYH10 antibody [EPR22564-23] - Goat IgG (Chimeric) - BSA and Azide free (ab300649)

Lane 1:

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

Lane 2:

HEK-293T (human embryonic kidney epithelial cell) whole cell lysate, with ab300649 at 1/30 dilution (2 μg in 0.35mg lysates)

Lane 3:

Goat monoclonal IgG instead of ab300649 in HEK-293T whole cell lysate

Secondary

All lanes:

Peroxidase conjugated Rabbit anti-Goat IgG (H+L) at 1/5000 dilution

Predicted band size: 229 kDa

Observed band size: 250 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-non-muscle Myosin IIB/MYH10 antibody [EPR22564-23] - Goat IgG (Chimeric) - BSA and Azide free (AB300649)

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling non-muscle Myosin IIB/MYH10 with ab300649 at 1/500 (1.96 ug/ml) followed by a Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution. Positive staining on mouse cerebellum. The section was incubated with ab300649 for 30 mins at room temperature, the 2nd antibody Donkey Anti-Goat IgG (HRP) ab205723 was used.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-non-muscle Myosin IIB/MYH10 antibody [EPR22564-23] - Goat IgG (Chimeric) - BSA and Azide free (AB300649)

Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labeling non-muscle Myosin IIB/MYH10 with ab300649 at 1/500 (1.96 ug/ml) followed by a Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution. Positive staining on rat cerebellum. The section was incubated with ab300649 for 30 mins at room temperature, the 2nd antibody Donkey Anti-Goat IgG (HRP) ab205723 was used.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is Donkey Anti-Goat IgG H&L (HRP) (ab205723) at 1 : 2000 dilution.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-non-muscle Myosin IIB/MYH10 antibody [EPR22564-23] - Goat IgG (Chimeric) - BSA and Azide free (AB300649)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling non-muscle Myosin IIB/MYH10 with ab300649 at 1/50 (19.56 ug/ml) dilution, followed by ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) antibody at 1/500 4ug/ml dilution (Green). Confocal image showing cytoskeletal staining in NIH/3T3 cell line is observed. 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 ab150129 Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) at 1/500 4ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-non-muscle Myosin IIB/MYH10 antibody [EPR22564-23] - Goat IgG (Chimeric) - BSA and Azide free (AB300649)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling non-muscle Myosin IIB/MYH10 with ab300649 at 1/1000 dilution (0.1ug) (Red) compared with a Goat IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Donkey anti-Goat IgG (Alexa Fluor® 488, ab150133) at 1/2000 dilution was used as the secondary antibody.

• WB

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Western blot - Anti-non-muscle Myosin IIB/MYH10 antibody [EPR22564-23] - Goat IgG (Chimeric) - BSA and Azide free (AB300649)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : Lane 1-2 : 10 seconds Lane 3-5 : 37 seconds

All lanes:

Western blot - Anti-non-muscle Myosin IIB/MYH10 antibody [EPR22564-23] - Goat IgG (Chimeric) - BSA and Azide free (ab300649) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

Human lung tissue lysate at 20 µg

Lane 4:

Mouse brain tissue lysate at 20 µg

Lane 5:

Rat brain tissue lysate at 20 µg

Secondary

All lanes:

Rabbit anti-Goat IgG (H+L), Peroxidase conjugated at 1/5000 dilution

Observed band size: 250 kDa

false

Exposure time: 10s