Anti-CYP51A1/CYP51 antibody [EPR27379-150]

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human testis tissue staining ATAD2 with ab324560 at a 1/100 dilution, ab323317 anti-Oligodendrocyte Specific Protein used at 1/500 dilution and ab315800 anti-CYP51A1/CYP51 used at a 1/2000 dilution.

Panel A : anti-ATAD2 (green; Opal™570), anti-Oligodendrocyte Specific Protein (magenta; Opal™690) and anti-CYP51A1/CYP51 (gray; Opal™520) on human testis.

Panel B : anti-ATAD2 staining nucleus of proliferating cells in human testis.

Panel C : anti-Oligodendrocyte Specific Protein staining apical Sertoli cell crypts in human testis.

Panel D : anti-CYP51A1/CYP51 staining Leydig cells in human testis.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324560, ab323317 and ab315800 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling CYP51A1/CYP51 with ab315800 at 1/2000 (0.26 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on leydig cells of human testis. The section was incubated with ab315800 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-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Huh7 (human hepatocellular carcinoma epithelial cell) cells labelling CYP51A1/CYP51 with ab315800 at 1/50 (10.38 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 cytoplasmic staining in Huh7 cell line.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

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 (2 ug/mL) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293 (human embryonic kidney epithelial cell) cells labelling CYP51A1/CYP51 with ab315800 at 1/50 dilution (1 ug)/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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human testis tissue staining Oligodendrocyte Specific Protein with ab323317 at a 1/500 dilution, ab315800 anti-CYP51A1 used at 1/2000 dilution and ab270534 anti-DDX4/MVH used at a 1/2000 dilution.

Panel A : anti-Oligodendrocyte Specific Protein (green; Opal™520), anti-CYP51A1 (magenta; Opal™690) and anti-DDX/MVH (gray; Opal™570) on human testis.

Panel B : anti-Oligodendrocyte Specific Protein staining apical Sertoli cell crypts in human testis.

Panel C : anti-CYP51A1 staining Leydig cells in human testis.

Panel D : anti-DDX4/MVH staining spermatogonia in human testis.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab323317, ab315800 and ab270534 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 Solution 2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling CYP51A1/CYP51 with ab315800 at 1/2000 (0.26 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human skeletal muscle. The section was incubated with ab315800 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-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human testis tissue staining ABHD2 with ab324946 at a 1/2000 dilution, ab315800 anti-CYP51A1 used at 1/2000 dilution ( 0.26 μg/ml) and ab290736 anti-IGF2BP1/IMP1 used at 1/2000 dilution ( 0.284 μg/ml).

Panel A : anti-ABHD2 (green; Opal™520), anti-CYP51A1 (magenta; Opal™690), anti-IGF2BP1/IMP1 (gray; Opal™570) on human testis.

Panel B : anti-ABHD2 staining spermatocytes in human testis.

Panel C : anti-CYP51A1 staining leydig cells in human testis.

Panel D : anti-IGF2BP1/IMP1 staining spermatogonia in mouse colon.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324946, ab315800 and ab290736 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-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human testis tissue staining Hormone sensitive lipase with ab325336 at a 1/2000 (0.248 μg/ml) dilution, ab315800 anti-CYP51A1 used at a 1/2000 (0.26 μg/ml) dilution, and ab270534 anti-DDX4 used at a 1/2000 (0.238 μg/ml) dilution.

Panel A : anti-Hormone sensitive lipase (green; Opal™520), anti-CYP51A1 (magenta; Opal™690), anti-DDX4 (gray; Opal™570) on human testis.

Panel B : anti-Hormone sensitive lipase staining spermatid in human testis.

Panel C : anti-CYP51A1 staining leydig cells in human testis.

Panel D : anti-DDX4 staining spermatogonia in human testis.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab325336, ab315800 and ab270534 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-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human testis tissue staining CYP51A1/CYP51 with ab315800 at a 1/2000 dilution, ab270534 anti-DDX4 / MVH used at 1/2000 dilution and ab315275 anti-LAMA1 used at a 1/500 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody.

Panel A : merged staining of anti-CYP51A1/CYP51 (green; Opal^™520), anti-DDX4 / MVH (magenta; Opal^™690) and anti-LAMA1 (grey; Opal^™570) on human testis. Panel B : anti-CYP51A1/CYP51 staining leydig cells in human testis.
Panel C : anti-DDX4 / MVH staining spermatogonia and spermatocytes in human testis.
Panel D : anti-LAMA1 staining basement membrane in human testis.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab315800, ab270834 and ab315275 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.

• IP

Supplier Data

Immunoprecipitation - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

CYP51A1/CYP51 was immunoprecipitated from 0.35 mg Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab315800 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315800 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (ab315800) at 1/1000 dilution

Lane 1:

Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate

Lane 2:

ab315800 at 1/30 IP in Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 1/30 dilution

Lane 3:

Rabbit monoclonal IgG (<a href='/ko/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab315800 in Huh7 whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/ko/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C6 (rat glial tumor glial cell) cells labelling CYP51A1/CYP51 with ab315800 at 1/500 dilution (0.1 ug)/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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling CYP51A1/CYP51 with ab315800 at 1/500 dilution (0.1 ug)/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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat testis tissue staining CYP51A1/CYP51 with ab315800 at a 1/2000 dilution, ab270534 anti-DDX4 / MVH used at 1/2000 dilution and ab315275 anti-LAMA1 used at a 1/500 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody.

Panel A : merged staining of anti-CYP51A1/CYP51 (green; Opal^™520), anti-DDX4 / MVH (magenta; Opal^™690) and anti-LAMA1 (grey; Opal^™570) on rat testis. Panel B : anti-CYP51A1/CYP51 staining leydig cells in rat testis.
Panel C : anti-DDX4 / MVH staining spermatogonia and spermatocytes in rat testis.
Panel D : anti-LAMA1 staining basement membrane in rat testis.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab315800, ab270834 and ab315275 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

Lab

Multiplex immunohistochemistry - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse testis tissue staining CYP51A1/CYP51 with ab315800 at a 1/2000 dilution, ab270534 anti-DDX4 / MVH used at 1/2000 dilution and ab315275 anti-LAMA1 used at a 1/500 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody.

Panel A : merged staining of anti-CYP51A1/CYP51 (green; Opal^™520), anti-DDX4 / MVH (magenta; Opal^™690) and anti-LAMA1 (grey; Opal^™570) on mouse testis. Panel B : anti-CYP51A1/CYP51 staining leydig cells in mouse testis.
Panel C : anti-DDX4 / MVH staining spermatogonia and spermatocytes in mouse testis.
Panel D : anti-LAMA1 staining basement membrane in mouse testis.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab315800, ab270834 and ab315275 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

Lab

Multiplex immunohistochemistry - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse testis tissue staining ATAD2 with ab324560 at a 1/500 dilution, ab323317 anti-Oligodendrocyte Specific Protein used at 1/500 dilution and ab315800 anti-CYP51A1/CYP51 used at a 1/2000 dilution.

Panel A : anti-ATAD2 (green; Opal™570), anti-Oligodendrocyte Specific Protein (magenta; Opal™690) and anti-CYP51A1/CYP51 (gray; Opal™520) on mouse testis.

Panel B : anti-ATAD2 staining nucleus of proliferating cells in mouse testis.

Panel C : anti-Oligodendrocyte Specific Protein staining apical Sertoli cell crypts in mouse testis.

Panel D : anti-CYP51A1/CYP51 staining Leydig cells in mouse testis.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324560, ab323317 and ab315800 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling CYP51A1/CYP51 with ab315800 at 1/2000 (0.26 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse skeletal muscle. The section was incubated with ab315800 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-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling CYP51A1/CYP51 with ab315800 at 1/2000 (0.26 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat skeletal muscle. The section was incubated with ab315800 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-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling CYP51A1/CYP51 with ab315800 at 1/2000 (0.26 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on leydig cells of rat testis. The section was incubated with ab315800 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-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling CYP51A1/CYP51 with ab315800 at 1/2000 (0.26 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on leydig cells of mouse testis. The section was incubated with ab315800 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-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat testis tissue staining Oligodendrocyte Specific Protein with ab323317 at a 1/500 dilution, ab315800 anti-CYP51A1 used at 1/2000 dilution and ab270534 anti-DDX4/MVH used at a 1/2000 dilution.

Panel A : anti-Oligodendrocyte Specific Protein (green; Opal™520), anti-CYP51A1 (magenta; Opal™690) and anti-DDX/MVH (gray; Opal™570) on rat testis.

Panel B : anti-Oligodendrocyte Specific Protein staining apical Sertoli cell crypts in rat testis.

Panel C : anti-CYP51A1 staining Leydig cells in rat testis.

Panel D : anti-DDX4/MVH staining spermatogonia in rat testis.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab323317, ab315800 and ab270534 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 Solution 2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse testis tissue staining Oligodendrocyte Specific Protein with ab323317 at a 1/500 dilution, ab315800 anti-CYP51A1 used at 1/2000 dilution and ab270534 anti-DDX4/MVH used at a 1/2000 dilution.

Panel A : anti-Oligodendrocyte Specific Protein (green; Opal™520), anti-CYP51A1 (magenta; Opal™690) and anti-DDX/MVH (gray; Opal™570) on human testis.

Panel B : anti-Oligodendrocyte Specific Protein staining apical Sertoli cell crypts in mouse testis.

Panel C : anti-CYP51A1 staining Leydig cells in mouse testis.

Panel D : anti-DDX4/MVH staining spermatogonia in mouse testis.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab323317, ab315800 and ab270534 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 Solution 2) for 20 mins.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C6 (rat glial tumor glial cell) cells labelling CYP51A1/CYP51 with ab315800 at 1/50 (10.38 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 cytoplasmic staining in C6 cell line.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

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 (2 ug/mL) dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling CYP51A1/CYP51 with ab315800 at 1/50 (10.38 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 cytoplasmic staining in NIH/3T3 cell line.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

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 (2 ug/mL) dilution.

• WB

Supplier Data

Western blot - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

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

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

All lanes:

Western blot - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (ab315800) at 1/1000 dilution

Lane 1:

HEK-293 (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HEK-293 transfected with siRNA specifically targeting CYP51A1 whole cell lysate at 20 µg

Lane 3:

Huh7 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 5:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 51 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (AB315800)

Negative control : skeletal muscle.

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

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

All lanes:

Western blot - Anti-CYP51A1/CYP51 antibody [EPR27379-150] (ab315800) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Human skeletal muscle tissue lysate at 20 µg

Lane 3:

Mouse liver tissue lysate at 20 µg

Lane 4:

Mouse testis tissue lysate at 20 µg

Lane 5:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 6:

Rat liver tissue lysate at 20 µg

Lane 7:

Rat testis tissue lysate at 20 µg

Lane 8:

Rat skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 51 kDa,36 kDa

false

Exposure time: 26s