Anti-ABCA4 antibody [EPR26543-66]

• IHC-FoFr

Lab

Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-ABCA4 antibody [EPR26543-66] (AB303504)

Immunocytochemistry/Immunofluorescence analysis of mouse retina cells labelling ABCA4 (Green) with ab303504 at 1/50 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Alexa Fluor^® 488 conguated Goat Anti-Rabbit IgG H&L (ab150081) preadsorbed, was used as the secondary antibody for the primary at a 1/1000 dilution. Anti-MAP2 mouse monoclonal antibody (ab11267) (Red) was used to counterstain at a 1/500 dilution, with Alexa Fluor^® 594 conguated Goat Anti-Mouse IgG H&L (ab150120) as the secondary (1/1000 dilution). DAPI (Blue) was used as the nuclear counterstain.

For negative (-ve) control 1, ab303504 (1/50 dilution) was used with secondary ab150120 (1/1000 dilution) and -ve control 2 used ab11267 (1/500 dilution) with secondary ab150081 (1/1000 dilution).

Confocal image showing cytoplasmic staining in mouse retina primary neuron.

Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection.

Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-ABCA4 antibody [EPR26543-66] (AB303504)

Immunocytochemistry/Immunofluorescence analysis of mouse retina cells labelling ABCA4 (Green) with ab303504 at 1/50 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Alexa Fluor^® 488 conguated Goat Anti-Rabbit IgG H&L (ab150081) preadsorbed, was used as the secondary antibody for the primary at a 1/1000 dilution. Anti-MAP2 mouse monoclonal antibody (ab11267) (Red) was used to counterstain at a 1/500 dilution, with Alexa Fluor^® 594 conguated Goat Anti-Mouse IgG H&L (ab150120) as the secondary (1/1000 dilution). DAPI (Blue) was used as the nuclear counterstain.

For negative (-ve) control 1, ab303504 (1/50 dilution) was used with secondary ab150120 (1/1000 dilution) and -ve control 2 used ab11267 (1/500 dilution) with secondary ab150081 (1/1000 dilution).

Confocal image showing cytoplasmic staining in mouse retina primary neuron.

Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection.

Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-ABCA4 antibody [EPR26543-66] (AB303504)

Immunocytochemistry/Immunofluorescence analysis of mouse neuron cells labelling ABCA4 (Green) with ab303504 at 1/50 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Alexa Fluor^® 488 conguated Goat Anti-Rabbit IgG H&L (ab150081) preadsorbed, was used as the secondary antibody for the primary at a 1/1000 dilution. Anti-MAP2 mouse monoclonal antibody (ab11267) (Red) was used to counterstain at a 1/500 dilution, with Alexa Fluor^® 594 conguated Goat Anti-Mouse IgG H&L (ab150120) as the secondary (1/1000 dilution). DAPI (Blue) was used as the nuclear counterstain.

For negative (-ve) control 1, ab303504 (1/50 dilution) was used with secondary ab150120 (1/1000 dilution) and -ve control 2 used ab11267 (1/500 dilution) with secondary ab150081 (1/1000 dilution).

Confocal image showing no staining in mouse primary neuron.

Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection.

Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-ABCA4 antibody [EPR26543-66] (AB303504)

Immunocytochemistry/Immunofluorescence analysis of rat retina cells labelling ABCA4 (Green) with ab303504 at 1/50 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Alexa Fluor^® 488 conguated Goat Anti-Rabbit IgG H&L (ab150081) preadsorbed, was used as the secondary antibody for the primary at a 1/1000 dilution. Anti-MAP2 mouse monoclonal antibody (ab11267) (Red) was used to counterstain at a 1/500 dilution, with Alexa Fluor^® 594 conguated Goat Anti-Mouse IgG H&L (ab150120) as the secondary (1/1000 dilution). DAPI (Blue) was used as the nuclear counterstain.

For negative (-ve) control 1, ab303504 (1/50 dilution) was used with secondary ab150120 (1/1000 dilution) and -ve control 2 used ab11267 (1/500 dilution) with secondary ab150081 (1/1000 dilution).

Confocal image showing cytoplasmic staining in rat retina primary neuron.

Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection.

Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-ABCA4 antibody [EPR26543-66] (AB303504)

Immunocytochemistry/Immunofluorescence analysis of rat neuron cells labelling ABCA4 (Green) with ab303504 at 1/50 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Alexa Fluor^® 488 conguated Goat Anti-Rabbit IgG H&L (ab150081) preadsorbed, was used as the secondary antibody for the primary at a 1/1000 dilution. Anti-MAP2 mouse monoclonal antibody (ab11267) (Red) was used to counterstain at a 1/500 dilution, with Alexa Fluor^® 594 conguated Goat Anti-Mouse IgG H&L (ab150120) as the secondary (1/1000 dilution). DAPI (Blue) was used as the nuclear counterstain.

For negative (-ve) control 1, ab303504 (1/50 dilution) was used with secondary ab150120 (1/1000 dilution) and -ve control 2 used ab11267 (1/500 dilution) with secondary ab150081 (1/1000 dilution).

Confocal image showing no staining in rat primary neuron.

Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection.

Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABCA4 antibody [EPR26543-66] (AB303504)

Immunohistochemical analysis of paraffin-embedded Mouse retina tissue labeling abCA4 with ab303504 at 1/500 (1.024 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on mouse retina. The section was incubated with ab303504 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-ABCA4 antibody [EPR26543-66] (AB303504)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling abCA4 with ab303504 at 1/500 (1.024 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control : no staining on mouse cardiac muscle. The section was incubated with ab303504 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-ABCA4 antibody [EPR26543-66] (AB303504)

Immunohistochemical analysis of paraffin-embedded Rat retina tissue labeling abCA4 with ab303504 at 1/500 (1.024 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on rat retina. The section was incubated with ab303504 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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-ABCA4 antibody [EPR26543-66] (AB303504)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat retina tissue staining RPE65 with ab231782 at a 1/8000 dilution, ab308463 anti-CPLX3 used at 1/5000 dilution and ab303504 anti-ABCA4 used at a 1/500 dilution.

Panel A : merged staining of anti-RPE65 (green; Opal™520), anti-CPLX3 (magenta; Opal™690) and anti-ABCA4 (gray; Opal™570) on rat retina.
Panel B : anti-RPE65 staining pigmented layer in rat retina.
Panel C : ant-CPLX3 staining ribbon synapses of photoreceptors and bipolar cells in rat retina.
Panel D : ant-ABCA4 staining photoreceptor outer segments in rat retina.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab231782, ab308463 and ab303504 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-ABCA4 antibody [EPR26543-66] (AB303504)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse retina tissue staining RPE65 with ab231782 at a 1/8000 dilution, ab308463 anti-CPLX3 used at 1/5000 dilution and ab303504 anti-ABCA4 used at a 1/500 dilution.

Panel A : merged staining of anti-RPE65 (green; Opal™520), anti-CPLX3 (magenta; Opal™690) and anti-ABCA4 (gray; Opal™570) on mouse retina.
Panel B : anti-RPE65 staining pigmented layer in mouse retina.
Panel C : ant-CPLX3 staining ribbon synapses of photoreceptors and bipolar cells in mouse retina.
Panel D : ant-ABCA4 staining photoreceptor outer segments in mouse retina.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab231782, ab308463 and ab303504 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Lab

Western blot - Anti-ABCA4 antibody [EPR26543-66] (AB303504)

Negative control : heart (PMID : 9054934).

Bands around 150 kDa may be due to degradation.

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) was used as a loading control staining.

All lanes:

Western blot - Anti-ABCA4 antibody [EPR26543-66] (ab303504) at 1/1000 dilution

Lane 1:

Mouse retina tissue lysate at 20 µg

Lane 2:

Mouse heart tissue lysate at 20 µg

Lane 3:

Rat eyeball tissue lysate at 20 µg

Lane 4:

Rat heart tissue 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 1/50000 dilution

Predicted band size: 256 kDa

Observed band size: 260 kDa

false

Exposure time: 59s