Anti-ADK antibody [EPR27497-67] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ADK antibody [EPR27497-67] - BSA and Azide free (AB307358)

This data was developed using ab307357, the same antibody clone in a different buffer formulation.  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized ADK KO HAP1 (ADK knockout human chronic myelogenous leukemia near-haploid cell) cells labeling ADK with ab307357 at 1/50 dilution (10.64 µg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 µg/ml) (Green). Confocal image showing nuclear and weak cytoplasmic staining in wildtype HAP1 cells, and no staining in ADK knockout HAP1 cells. The 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 dilution (2.5 µg/ml) (Red). 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 dilution (2 µg/ml).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ADK antibody [EPR27497-67] - BSA and Azide free (AB307358)

This data was developed using ab307357, the same antibody clone in a different buffer formulation.  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) (Right) / ADK knockout HAP1 (Left) cells labeling ADK with ab307357 at 1/50 dilution (1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-ADK antibody [EPR27497-67] - BSA and Azide free (AB307358)

This data was developed using ab307357, the same antibody clone in a different buffer formulation.  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling ADK with ab307357 at 1/50 dilution (1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ADK antibody [EPR27497-67] - BSA and Azide free (AB307358)

This data was developed using ab307357, the same antibody clone in a different buffer formulation.  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling ADK with ab307357 at 1/50 dilution (10.64 µg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 µg/ml) (Green). Confocal image showing nuclear and weak cytoplasmic staining in NIH/3T3 cells. The 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 dilution (2.5 µg/ml) (Red). 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 dilution (2 µg/ml).

• IP

Supplier Data

Immunoprecipitation - Anti-ADK antibody [EPR27497-67] - BSA and Azide free (AB307358)

This data was developed using ab307357, the same antibody clone in a different buffer formulation. ADK was immunoprecipitated from 0.35 mg RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 µg with ab307357 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307357 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : RAW264.7 whole cell lysate 10 µg Lane 2 : ab307357 IP in RAW264.7 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307357 in RAW264.7 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 128 seconds

All lanes:

Immunoprecipitation - Anti-ADK antibody [EPR27497-67] (<a href='/en-us/products/primary-antibodies/adk-antibody-epr27497-67-ab307357'>ab307357</a>) at 1/1000 dilution

Lane 1:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

Lane 2:

RAW264.7 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

Observed band size: 45 kDa

false

Exposure time: 128s

• IP

Supplier Data

Immunoprecipitation - Anti-ADK antibody [EPR27497-67] - BSA and Azide free (AB307358)

This data was developed using ab307357, the same antibody clone in a different buffer formulation. ADK was immunoprecipitated from 0.35 mg C6 (rat glial tumor glial cell) whole cell lysate 10 µg with ab307357 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307357 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : C6 whole cell lysate 10 µg Lane 2 : ab307357 IP in C6 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307357 in C6 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 128 seconds.

All lanes:

Immunoprecipitation - Anti-ADK antibody [EPR27497-67] (<a href='/en-us/products/primary-antibodies/adk-antibody-epr27497-67-ab307357'>ab307357</a>) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell) whole cell lysate

Lane 2:

C6 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

Observed band size: 45 kDa

false

Exposure time: 128s

• WB

Supplier Data

Western blot - Anti-ADK antibody [EPR27497-67] - BSA and Azide free (AB307358)

This data was developed using 307357, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : Low expression : A549 In Western blot, ab307357 was shown to bind specifically to ADK. A band was observed at 45 kDa in wild-type HAP1 cell lysates whereas no signal observed at this size in ADK knockout cell line. This blot (lanes 1-4) was developed using a high sensitivity ECL substrate. 125 seconds Exposure time :

All lanes:

Western blot - Anti-ADK antibody [EPR27497-67] (<a href='/en-us/products/primary-antibodies/adk-antibody-epr27497-67-ab307357'>ab307357</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate

Lane 2:

ADK knockout HAP1 whole cell lysate

Lane 3:

HeLa whole cell lysate

Lane 4:

A549 whole cell lysate

Lane 5:

RAW264.7 whole cell lysate

Lane 6:

C6 whole cell lysate

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: 45 kDa

true

Exposure time: 125s

• WB

Supplier Data

Western blot - Anti-ADK antibody [EPR27497-67] - BSA and Azide free (AB307358)

This data was developed using 307357, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : mouse heart (PMID : 15930047;14736855), rat heart (PMID : 15930047;14736855). Exposure time : 15 seconds

All lanes:

Western blot - Anti-ADK antibody [EPR27497-67] (<a href='/en-us/products/primary-antibodies/adk-antibody-epr27497-67-ab307357'>ab307357</a>) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate

Lane 2:

Rat liver tissue lysate

Lane 3:

Mouse heart tissue lysate

Lane 4:

Rat heart tissue lysate

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: 45 kDa

false

Exposure time: 15s