Anti-DCK antibody [EPR29105-53]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DCK antibody [EPR29105-53] (AB317829)

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling DCK with ab317829 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on human cardiac muscle. The section was incubated with ab317829 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-DCK antibody [EPR29105-53] (AB317829)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling DCK with ab317829 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil (PMID : 29550884). The section was incubated with ab317829 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

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-DCK antibody [EPR29105-53] (AB317829)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) (Right) / IMR-90 (human lung fibroblast) (Left) cells labelling DCK with ab317829 at 1/500 dilution (0.1ug) (Red) (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.

Low expression : IMR-90 (PMID : 33556172).

• IP

Supplier Data

Immunoprecipitation - Anti-DCK antibody [EPR29105-53] (AB317829)

DCK was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab317829 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317829 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab317829 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317829 in HeLa whole cell lysate

All lanes:

Immunoprecipitation - Anti-DCK antibody [EPR29105-53] (ab317829) at 1/30 dilution

All lanes:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

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

false

Exposure time: 180s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DCK antibody [EPR29105-53] (AB317829)

Immunohistochemical analysis of paraffin-embedded Mouse thymus tissue labeling DCK with ab317829 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse thymus. The section was incubated with ab317829 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-DCK antibody [EPR29105-53] (AB317829)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling DCK with ab317829 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : scattered positive staining on mouse cardiac muscle. The section was incubated with ab317829 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-DCK antibody [EPR29105-53] (AB317829)

Immunohistochemical analysis of paraffin-embedded Mouse T-cell lymphoma tissue labeling DCK with ab317829 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse T-cell lymphoma. The section was incubated with ab317829 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-DCK antibody [EPR29105-53] (AB317829)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling DCK with ab317829 at 1/500 (1.008 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen. The section was incubated with ab317829 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

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Immunocytochemistry/ Immunofluorescence - Anti-DCK antibody [EPR29105-53] (AB317829)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling DCK with ab317829 at 1/500 (1.008 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 nuclear staining in RAW 264.7 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Low expression : A20.
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 (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.

• IP

Supplier Data

Immunoprecipitation - Anti-DCK antibody [EPR29105-53] (AB317829)

DCK was immunoprecipitated from 0.35 mg RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate with ab317829 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317829 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 2 : ab317829 IP in RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317829 in RAW 264.7 whole cell lysate

All lanes:

Immunoprecipitation - Anti-DCK antibody [EPR29105-53] (ab317829) at 1/30 dilution

All lanes:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

Secondary

All lanes:

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

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-DCK antibody [EPR29105-53] (AB317829)

Low expression : IMR-90 (PMID : 33556172), A20.

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

All lanes:

Western blot - Anti-DCK antibody [EPR29105-53] (ab317829) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) whole cell lysate at 30 µg

Lane 2:

HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 30 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 30 µg

Lane 4:

IMR-90 (human lung fibroblast) whole cell lysate at 30 µg

Lane 5:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 30 µg

Lane 6:

A20 (mouse reticulum sarcoma b lymphocyte) whole cell lysate at 30 µg

Lane 7:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 30 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: 31 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-DCK antibody [EPR29105-53] (AB317829)

Low expression : liver (PMID : 7929097, PMID : 9079672), placenta (PMID : 9079672).

The identity of the bands higher than 25 kDa (in lane 2) are unknown.

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-DCK antibody [EPR29105-53] (ab317829) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 30 µg

Lane 2:

Human placenta tissue lysate at 30 µg

Lane 3:

Human thymus tissue lysate at 30 µg

Lane 4:

Human liver tissue lysate at 30 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 31 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-DCK antibody [EPR29105-53] (AB317829)

Low expression : liver, heart (PMID : 7929097).

The identity of the higher MW band at approximately 40 kDa (in lane 2) is unknown.

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-DCK antibody [EPR29105-53] (ab317829) at 1/1000 dilution

Lane 1:

Mouse thymus tissue lysate at 30 µg

Lane 2:

Mouse liver tissue lysate at 30 µg

Lane 3:

Mouse heart tissue lysate at 30 µg

Lane 4:

Mouse spleen tissue lysate at 30 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: 31 kDa,36 kDa

false

Exposure time: 59s