Anti-MafA antibody [EPR30417-538] - BSA and Azide free

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-MafA antibody [EPR30417-538] - BSA and Azide free (AB324879)

This data was developed using ab324869, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse pancreas tissue staining MafA with ab324869 at a 1 : 100 (5.13 ug/ml) dilution, ab181547 anti-Insulin used at 1 : 20000 (0.053 ug/ml) dilution and ab209792 anti-GIP used at a 1 : 2000 (0.26 ug/ml) dilution.

Panel A : merged staining of anti-MafA (green; Opal™520), anti-Insulin (magenta; Opal™690) and anti-GIP (gray; Opal™570) on mouse pancreas.
Panel B : anti-MafA staining nucleus of beta cells in mouse pancreas islet.
Panel C : anti-Insulin staining cytoplasm of beta cells in mouse pancreas islet.
Panel D : anti-GIP staining the alpha cells in mouse pancreas islet.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324869, ab181547 and ab209792 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-MafA antibody [EPR30417-538] - BSA and Azide free (AB324879)

This data was developed using ab324869, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat pancreas tissue staining MafA with ab324869 at a 1 : 100 (5.13 ug/ml) dilution, ab181547 anti-Insulin used at 1 : 20000 (0.053 ug/ml) dilution and ab209792 anti-GIP used at a 1 : 2000 (0.26 ug/ml) dilution.

Panel A : merged staining of anti-MafA (green; Opal™520), anti-Insulin (magenta; Opal™690) and anti-GIP (gray; Opal™570) on rat pancreas.
Panel B : anti-MafA staining nucleus of beta cells in rat pancreas islet.
Panel C : anti-Insulin staining cytoplasm of beta cells in rat pancreas islet.
Panel D : anti-GIP staining the alpha cells in rat pancreas islet.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324869, ab181547 and ab209792 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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-MafA antibody [EPR30417-538] - BSA and Azide free (AB324879)

This data was developed using ab324869, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling MafA with ab324869 at 1/100 (5.13 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on rat cerebrum.
The primary antibody was incubated 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 was performed 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-MafA antibody [EPR30417-538] - BSA and Azide free (AB324879)

This data was developed using ab324869, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling MafA with ab324869 at 1/100 (5.13 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse cerebrum (PMID : 12011435).
The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MafA antibody [EPR30417-538] - BSA and Azide free (AB324879)

This data was developed using ab324869, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Beta-TC-6 (mouse pancreas insulinoma beta cell) cells labelling MafA with ab324869 at 1/50 (10.26 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing nuclear staining in Beta-TC-6 cells (shown in green). The counterstain was observed in magenta.
Low expression : C2C12.
Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 (1 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2 ug/ml) dilution (Magenta).

-ve control 1 : ab324869 at a 1/50 dilution followed by ab150120 at a 1/1000 dilution.

-ve control 2 : ab7291 at a 1/1000 dilution followed by ab150081 at a 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MafA antibody [EPR30417-538] - BSA and Azide free (AB324879)

This data was developed using ab324869, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling MafA with ab324869 at 1/100 (5.13 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on islets of mouse pancreas (PMID : 12917329).
The primary antibody was incubated 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 was performed 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-MafA antibody [EPR30417-538] - BSA and Azide free (AB324879)

This data was developed using ab324869, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat pancreas tissue labeling MafA with ab324869 at 1/100 (5.13 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on islets of rat pancreas.
The primary antibody was incubated 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-MafA antibody [EPR30417-538] - BSA and Azide free (AB324879)

This data was developed using ab324869, the same antibody clone in a different buffer formulation.

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

Low expression : C2C12

Negative control : NIH/3T3

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

All lanes:

Western blot - Anti-MafA antibody [EPR30417-538] (<a href='/en-us/products/primary-antibodies/mafa-antibody-epr30417-538-ab324869'>ab324869</a>) at 1/1000 dilution

Lane 1:

Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate at 40 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 40 µg

Lane 3:

C2C12 (mouse myoblast) whole cell lysate at 40 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 50 kDa,36 kDa

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-MafA antibody [EPR30417-538] - BSA and Azide free (AB324879)

This data was developed using ab324869, the same antibody clone in a different buffer formulation.

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

Negative control : brain (PMID : 12011435)

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

All lanes:

Western blot - Anti-MafA antibody [EPR30417-538] (<a href='/en-us/products/primary-antibodies/mafa-antibody-epr30417-538-ab324869'>ab324869</a>) at 1/1000 dilution

Lane 1:

Mouse pancreas tissue lysate at 40 µg

Lane 2:

Mouse brain tissue lysate at 40 µg

Lane 3:

Rat pancreas tissue lysate at 40 µg

Lane 4:

Rat brain 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: 50 kDa,36 kDa

false

Exposure time: 92s