Anti-Arx antibody [EPR27481-32]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Arx antibody [EPR27481-32] (AB308260)

Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling Arx with ab308260 at 1/50 dilution (10.35 ug/ml) followed by ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human pancreas islet (PMID : 31263286). The section was incubated with ab308260 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Arx antibody [EPR27481-32] (AB308260)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Arx with ab308260 at 1/50 dilution (10.35 ug/ml) followed by ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Negative control : no staining on human kidney (PMID : 12359145). The section was incubated with ab308260 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

• mIHC

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Multiplex immunohistochemistry - Anti-Arx antibody [EPR27481-32] (AB308260)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human pancreas tissue staining Arx with ab308260 at a 1/50 dilution ( 10.35 μg/ml) and ab271989 anti-GIP used at a 1/4000 dilution (0.27 μg/ml).

Panel A : merged staining of anti-GIP (green; Opal™520) and anti-Homeobox protein ARX (red; Opal™570) on human pancreas.
Panel B : anti-Homeobox protein ARX stained on alpha cells.
Panel C : anti-GIP stained on alpha cells.

The section was incubated in two rounds of staining : in the order of ab308260 and ab271989 for 30 mins at room temperature.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. 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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Arx antibody [EPR27481-32] (AB308260)

Immunohistochemical analysis of paraffin-embedded rat E11.5 brain tissue labeling Arx with ab308260 at 1/50 dilution (10.35 ug/ml) followed by ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on rat E11.5 brain (PMID : 11971879). The section was incubated with ab308260 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Arx antibody [EPR27481-32] (AB308260)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized rat primary neural/glia cells labelling Arx with ab308260 at 1/100 dilution (5.175 ug/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green). Confocal image showing nuclear staining in rat primary neural/glia cell. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 dilution (4 ug/ml) followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (2 ug/ml) (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 dilution (2 ug/ml).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Arx antibody [EPR27481-32] (AB308260)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse primary neural/glia cells labelling Arx with ab308260 at 1/100 dilution (5.175 ug/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green). Confocal image showing nuclear staining in mouse primary neural/glia cell. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 dilution (4 ug/ml) followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (2 ug/ml) (Red). The nuclear counterstain was DAPI (Blue). -ve control 1 : ab308260 at 1/100 dilution followed by ab150120 at 1/1000 dilution. -ve control 2 : ab11267 at 1/500 dilution followed by ab150081 at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Arx antibody [EPR27481-32] (AB308260)

Immunohistochemical analysis of paraffin-embedded mouse E10.5 brain tissue labeling Arx with ab308260 at 1/50 dilution (10.35 ug/ml) followed by ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on mouse E10.5 brain (PMID : 11971879). The section was incubated with ab308260 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Arx antibody [EPR27481-32] (AB308260)

Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue labeling Arx with ab308260 at 1/50 dilution (10.35 ug/ml) followed by ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on mouse pancreas islet. The section was incubated with ab308260 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Arx antibody [EPR27481-32] (AB308260)

Immunohistochemical analysis of paraffin-embedded rat pancreas tissue labeling Arx with ab308260 at 1/50 dilution (10.35 ug/ml) followed by ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on rat pancreas islet. The section was incubated with ab308260 at 4°C overnight. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Arx antibody [EPR27481-32] (AB308260)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized mouse primary neuron cells labelling Arx with ab308260 at 1/50 dilution (1ug) (Right panel) compared with a Rabbit monoclonal IgG isotype control (ab172730) (Left panel). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Cells were stained with rabbit IgG or ab308260.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Arx antibody [EPR27481-32] (AB308260)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized rat primary neuron cells labelling Arx with ab308260 at 1/50 dilution (1ug) (Right panel) compared with a Rabbit monoclonal IgG isotype control (ab172730) (Left panel). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Cells were stained with rabbit IgG or ab308260.

• WB

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Western blot - Anti-Arx antibody [EPR27481-32] (AB308260)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : liver(PMID : 12359145). Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-Arx antibody [EPR27481-32] (ab308260) at 1/1000 dilution

Lane 1:

Rat E12.5 brain lysate at 20 µg

Lane 2:

Rat E12.5 liver 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: 75 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Arx antibody [EPR27481-32] (AB308260)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : liver (PMID : 12359145). Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds.

All lanes:

Western blot - Anti-Arx antibody [EPR27481-32] (ab308260) at 1/1000 dilution

Lane 1:

Mouse E12.5 brain lysate at 20 µg

Lane 2:

Mouse E12.5 liver 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

Observed band size: 75 kDa

false