Anti-PCGF6 antibody [EPR26301-72]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PCGF6 antibody [EPR26301-72] (AB300059)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293T (Human embryonic kidney epithelial cell) cells labelling PCGF6 with ab300059 at 1/50 dilution (1μg) (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). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PCGF6 antibody [EPR26301-72] (AB300059)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeilized 293T (human embryonic kidney epithelial cell) cells lebelling PCGF6 with ab300059 at 1/500 (0.914 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 2 μg/mL dilution (Green). Confocal image showing nuclear staining in 293T cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 2.5 μg/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 μg/mL dilution.

• IP

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Immunoprecipitation - Anti-PCGF6 antibody [EPR26301-72] (AB300059)

PCGF6 was immunoprecipitated from 0.35 mg HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 10 μg with ab300059 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300059 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 10 μg

Lane 2 : ab300059 in HEK-293 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab300059 in HEK-293 whole cell lysate

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

Exposure time : 7.75 seconds

All lanes:

Immunoprecipitation - Anti-PCGF6 antibody [EPR26301-72] (ab300059)

Predicted band size: 39 kDa

false

• WB

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Western blot - Anti-PCGF6 antibody [EPR26301-72] (AB300059)

5% NFDM/TBST was used as blocking and diluting buffer and concentration.

The expression profile/molecular weight observed is consistent with that described in the literature (PMID : 28049731).
This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Western blot - Anti-PCGF6 antibody [EPR26301-72] (ab300059) at 1000 Cells

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell 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

Predicted band size: 39 kDa

Observed band size: 48 kDa,50 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-PCGF6 antibody [EPR26301-72] (AB300059)

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

Exposure time :
Lane 1 : 3 minutes
Lane 2 : 26 seconds

The expression profile/molecular weight observed is consistent with that described in the literature (PMID : 28049731).

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-PCGF6 antibody [EPR26301-72] (ab300059) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Human thymus tissue lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 39 kDa

Observed band size: 48 kDa,50 kDa

true