Anti-DYNC1H1 antibody [EPR30299-93]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-DYNC1H1 antibody [EPR30299-93] (AB325710)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA and HeLa transfected with siRNA targeting DYNC1H1 cells labelling DYNC1H1 with ab325710 at 1/500 (1.062 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in HeLa cells (shown in green) transfected with scrambled siRNA, showing decreased staining in HeLa cells transfected siRNA targeting DYNC1H1. The counterstain was observed in magenta. 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 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DYNC1H1 antibody [EPR30299-93] (AB325710)

Immunohistochemical analysis of paraffin-embedded Human astrocytoma tissue labeling DYNC1H1 with ab325710 at 1/5000 (0.106 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human astrocytoma.

The primary antibody was incubated for 10 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 10 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DYNC1H1 antibody [EPR30299-93] (AB325710)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling DYNC1H1 with ab325710 at 1/5000 (0.106 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human testis.

The primary antibody was incubated for 10 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 10 mins

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-DYNC1H1 antibody [EPR30299-93] (AB325710)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling DYNC1H1 with ab325710 at 1/500 (1.062 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in HeLa cell line (shown in green). The counterstain was observed in magenta. 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 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab325710 at 1/500 dilution, followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-DYNC1H1 antibody [EPR30299-93] (AB325710)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling DYNC1H1 with ab325710 at 1/500 (1.062 µg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in Neuro-2a cell line (shown in green). The counterstain was observed in magenta. 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 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab325710 at 1/500 dilution, followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 at 1/1000 dilution.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-DYNC1H1 antibody [EPR30299-93] (AB325710)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (fresh frozen) tissue labeling DYNC1H1 with ab325710 at 1/200 (2.655 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green).

Low expression : confocal image showing no staining on mouse liver. The nuclear counterstain was DAPI (Blue). The section was incubated with ab325710 for 60 mins at room temperature. 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).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-DYNC1H1 antibody [EPR30299-93] (AB325710)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum (fresh frozen) tissue labeling DYNC1H1 with ab325710 at 1/200 (2.655 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green).

Panel A : merged staining of anti-DYNC1H1 (ab325710, green), anti-NeuN (ab190565, magenta) on mouse cerebrum.
Panel B : anti-DYNC1H1 stained on mouse cerebrum.
Panel C : anti-NeuN stained in neurons of mouse cerebrum.
The section was incubated in two rounds of staining : in the order of ab325710 and ab190565, for 1 hr at room temperature. The nuclear counterstain was DAPI (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).

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-DYNC1H1 antibody [EPR30299-93] (AB325710)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse testis (fresh frozen) tissue labeling DYNC1H1 with ab325710 at 1/200 (2.655 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green).

Panel A : merged staining of anti-DYNC1H1 (ab325710, green), anti-SPC3 (ab97672, magenta) on mouse testis.
Panel B : anti-DYNC1H1 stained on mouse testis.
Panel C : anti-SPC3 stained on mouse testis.
The section was incubated with ab325710 and ab97672 for 1 hr at room temperature. The nuclear counterstain was DAPI (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).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-DYNC1H1 antibody [EPR30299-93] (AB325710)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling DYNC1H1 with ab325710 at 1/500 (1.062 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in mouse primary neural/glia cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). 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 (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab325710 at 1/500 dilution, followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab11267 at 1/500 dilution, followed by ab150081 at 1/1000 dilution.

• WB

Lab

Western blot - Anti-DYNC1H1 antibody [EPR30299-93] (AB325710)

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

Low expression : liver.

The bands beneath the target band (532 kDa) are likely to be degraded target fragments.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-DYNC1H1 antibody [EPR30299-93] (ab325710) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

Hela transfected with siRNA specifically targeting DYNC1H1 whole cell lysate at 20 µg

Lane 3:

Human cerebellum tissue lysate at 20 µg

Lane 4:

Human testis tissue lysate at 20 µg

Lane 5:

Human liver 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 1/100000 dilution

Observed band size: 532 kDa,36 kDa

false

Exposure time: 15s