Anti-MiTF antibody [EPR26363-10] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MiTF antibody [EPR26363-10] - BSA and Azide free (AB303531)

This data was developed using ab303530, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SK-MEL-28 (human malignant melanoma cell) cells labelling MiTF with ab303530 at 1/50 (11.32 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 mostly nuclear staining in SK-MEL-28 cell line.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). 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 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-MiTF antibody [EPR26363-10] - BSA and Azide free (AB303531)

This data was developed using ab303530, the same antibody clone in a different buffer formulation.Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SK-MEL-28 (human malignant melanoma) cells labelling MiTF with ab303530 at 1/500 dilution (0.1ug) (Red) (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 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MiTF antibody [EPR26363-10] - BSA and Azide free (AB303531)

This data was developed using ab303530, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human melanoma tissue labeling MiTF with ab303530 at 1/5000 (0.113 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human melanoma. The section was incubated with ab303530 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 LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MiTF antibody [EPR26363-10] - BSA and Azide free (AB303531)

This data was developed using ab303530, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling MITF with ab303530 at 1/100 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081).

The primary antibody was incubated for 60 mins at room temperature (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount^®. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 40 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MiTF antibody [EPR26363-10] - BSA and Azide free (AB303531)

This data was developed using ab303530, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human hair follicle tissue labeling MITF with ab303530 at 1/100 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081).

The primary antibody was incubated for 60 mins at room temperature (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Fluoromount^®. The immunostaining was performed on a Leica Biosystems BOND RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150081).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 40 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MiTF antibody [EPR26363-10] - BSA and Azide free (AB303531)

This data was developed using ab303530, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Mouse embryonic eye tissue labeling MiTF with ab303530 at 1/5000 (0.113 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse embryonic eye. The section was incubated with ab303530 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 LeicaDS9800 (Bond™ Polymer Refine Detection).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-MiTF antibody [EPR26363-10] - BSA and Azide free (AB303531)

This data was developed using ab303530, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Rat embryonic eye tissue labeling MiTF with ab303530 at 1/5000 (0.113 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on rat embryonic eye. The section was incubated with ab303530 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 LeicaDS9800 (Bond™ Polymer Refine Detection).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-MiTF antibody [EPR26363-10] - BSA and Azide free (AB303531)

This data was developed using ab303530, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Mouse skin tissue labeling MiTF with ab303530 at 1/5000 (0.113 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse skin. The section was incubated with ab303530 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 LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MiTF antibody [EPR26363-10] - BSA and Azide free (AB303531)

This data was developed using ab303530, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized B16-F10 (mouse skin melanoma cell) cells labelling MiTF with ab303530 at 1/50 (11.32 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 mostly nuclear staining in B16-F0 cell line.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). 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 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-MiTF antibody [EPR26363-10] - BSA and Azide free (AB303531)

This data was developed using ab303530, the same antibody clone in a different buffer formulation.Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized B16F0 (mouse melanoma epithelial cell-like) cells labelling MiTF with ab303530 at 1/500 dilution (0.1ug) (Red) (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 1/2000 dilution was used as the secondary antibody.

• WB

Supplier Data

Western blot - Anti-MiTF antibody [EPR26363-10] - BSA and Azide free (AB303531)

This data was developed using ab303530, the same antibody clone in a different buffer formulation.

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

Exposure time :

Lanes 1, 3, 4 : 70 seconds

Lane 2 : 180 seconds

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 17182576).

All lanes:

Western blot - Anti-MiTF antibody [EPR26363-10] (<a href='/en-us/products/primary-antibodies/mitf-antibody-epr26363-10-ab303530'>ab303530</a>) at 1/1000 dilution

Lane 1:

SK-MEL-28 (human malignant melanoma), whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

B16F0 (mouse melanoma epithelial cell-like), whole cell lysate at 20 µg

Lane 4:

B16F10 (mouse skin melanoma 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 1/100000 dilution

Predicted band size: 58 kDa

Observed band size: 40-70 kDa

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