Anti-DKKL1 antibody [EPR25721-68]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-DKKL1 antibody [EPR25721-68] (AB316308)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling DKKL1 with ab316308 at 1/100 (5.14 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 positive staining in 293T cells transfected with a human DKKL1 expression vector containing a myc-His-tag®. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain at 1/100 (5ug/ml) dilution (Magenta). 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-DKKL1 antibody [EPR25721-68] (AB316308)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T cells transfected with a human DKKL1 expression vector containing a myc-His-tag® (Middle) / 293T cells transfected with an empty expression vector containing a myc-His-tag® (Right) cells labelling DKKL1 with ab316308 at 1/500 dilution (0.1ug)/ Middle and Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

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

Cells are co-stained with Myc tag conjugated to Alexa Fluor®647.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-DKKL1 antibody [EPR25721-68] (AB316308)

Multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded Human testis tissue sections labeling DKKL1 with ab316308 at a 1/2000 dilutions (B), Wilms Tumor Protein with ab275970 at a 1/1200 dilutions (C), and SAGE1 with ab233388 at a 1/250 dilutions (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody.

Panel A : merged staining of anti-DKKL1 (red; Opal™ 570), anti-Wilms Tumor Protein (green; Opal™ 520) and anti-SAGE1 (gray; Opal™ 690) on human testis.
Panel B : anti-DKKL1 staining mature sperm cells in human testis.
Panel C : anti-Wilms Tumor Protein staining sertoli cells in human testis.
Panel D : anti-SAGE1 staining spermatogonia in human testis.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. The section was incubated in three rounds of staining : in the order of ab316308, ab275970, and ab233388 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

DAPI was used as a nuclear counterstain.

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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DKKL1 antibody [EPR25721-68] (AB316308)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling DKKL1 with ab316308 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human testis (PMID : 27699219).
The section was incubated with ab316308 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-DKKL1 antibody [EPR25721-68] (AB316308)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling DKKL1 with ab316308 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human cerebrum.
The section was incubated with ab316308for 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-DKKL1 antibody [EPR25721-68] (AB316308)

Immunohistochemistry analysis of Formalin/PFA-fixed paraffin-embedded sections Rat testis tissue labelling Lin28A with ab279647 at 1 : 1000 dilution (B), Wilms Tumor Protein with ab267377 at 1 : 500 dilution (C) and DKKL1 with ab316308 at 1 : 2000 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Lin28A (green; Opal™520), anti-Wilms Tumor Protein (magenta; Opal™690) and anti-DKKL1 (gray; Opal™570) on rat testis.

Panel B : anti-Lin28A staining undifferentiated spermatogonia in rat testis.

Panel C : ant-Wilms Tumor Protein staining Sertoli cells in rat testis.

Panel D : ant-DKKL11 staining mature sperm cells 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 ab279647, ab267377 and ab316308 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DKKL1 antibody [EPR25721-68] (AB316308)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling DKKL1 with ab316308 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse cerebrum (PMID : 15892050).
The section was incubated with ab316308 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-DKKL1 antibody [EPR25721-68] (AB316308)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling DKKL1 with ab316308 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat cerebrum (PMID : 15892050).
The section was incubated with ab316308 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-DKKL1 antibody [EPR25721-68] (AB316308)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling DKKL1 with ab316308 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse testis (PMID : 22817830).
The section was incubated with ab316308 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-DKKL1 antibody [EPR25721-68] (AB316308)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling DKKL1 with ab316308 at 1/2000 (0.257 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat testis.
The section was incubated with ab316308 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-DKKL1 antibody [EPR25721-68] (AB316308)

Immunohistochemistry analysis of Formalin/PFA-fixed paraffin-embedded sections Mouse testis tissue labelling Lin28A with ab279647 at 1 : 1000 dilution (B), Wilms Tumor Protein with ab267377 at 1 : 500 dilution (C) and DKKL1 with ab316308 at 1 : 2000 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-Lin28A (green; Opal™520), anti-Wilms Tumor Protein (magenta; Opal™690) and anti-DKKL1 (gray; Opal™570) on rat testis.

Panel B : anti-Lin28A staining undifferentiated spermatogonia in rat testis.

Panel C : ant-Wilms Tumor Protein staining Sertoli cells in rat testis.

Panel D : ant-DKKL11 staining mature sperm cells 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 ab279647, ab267377 and ab316308 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.

• WB

Supplier Data

Western blot - Anti-DKKL1 antibody [EPR25721-68] (AB316308)

Negative control : lung (PMID : 15892050), brain (PMID : 15892050).

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

All lanes:

Western blot - Anti-DKKL1 antibody [EPR25721-68] (ab316308) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 40 µg

Lane 2:

Human lung tissue lysate at 40 µg

Lane 3:

Human brain tissue 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: 32 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-DKKL1 antibody [EPR25721-68] (AB316308)

Negative control : brain (PMID : 15892050).

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

All lanes:

Western blot - Anti-DKKL1 antibody [EPR25721-68] (ab316308) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 40 µg

Lane 2:

Mouse 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: 32 kDa,36 kDa

false

Exposure time: 180s