重组Anti-VKORC1抗体[EPR20245]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VKORC1 antibody [EPR20245] (AB206656)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling VKORC1 with ab206656 at 1/100 dilution followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HepG2 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-VKORC1 antibody [EPR20245] (AB206656)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized A549 (Human lung carcinoma cell line) cells labeling VKORC1 with ab206656 at 1/100 dilution followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on A549 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-VKORC1 antibody [EPR20245] (AB206656)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling VKORC1 with ab206656 at 1/60 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluorr^® 488) at 1/2000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-VKORC1 antibody [EPR20245] (AB206656)

VKORC1 was immunoprecipitated from 0.35 mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab206656 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab206656 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : HepG2 whole cell lysate, 10 μg (Input).

Lane 2 : ab206656 IP in HepG2 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab206656 in HepG2 whole cell lysate.

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

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-VKORC1 antibody [EPR20245] (ab206656)

Predicted band size: 18 kDa

Observed band size: 15 kDa

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• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VKORC1 antibody [EPR20245] (AB206656)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling VKORC1 with ab206656 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on hepatocytes of mouse liver is observed [PMID : 25747820] [PMID : 14765194]. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-VKORC1 antibody [EPR20245] (AB206656)

Western blot : Anti-VKORC1 antibody [EPR20245] ab206656 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin ab7291 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 15 kDa in Wild-type HEK-293T cell lysates with no signal observed at this size in VKORC1 knockout HEK-293T cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-VKORC1 antibody [EPR20245] (ab206656) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysates at 20 µg

Lane 2:

Western blot - Human VKORC1 knockout HEK-293T cell line (<a href='/products/cell-lines/human-vkorc1-knockout-hek-293t-cell-line-ab266095'>ab266095</a>) at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Lane 4:

MCF7 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 18 kDa

Observed band size: 15 kDa

false

• WB

Supplier Data

Western blot - Anti-VKORC1 antibody [EPR20245] (AB206656)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-VKORC1 antibody [EPR20245] (ab206656) at 1/1000 dilution

Lane 1:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 3:

Mouse liver lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 18 kDa

Observed band size: 15 kDa

false

Exposure time: 8s

• WB

Unknown

Western blot - Anti-VKORC1 antibody [EPR20245] (AB206656)

Blocking/Dilution buffer : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 25753038).

All lanes:

Western blot - Anti-VKORC1 antibody [EPR20245] (ab206656) at 1/1000 dilution

Lane 1:

Human fetal kidney lysate at 10 µg

Lane 2:

Human fetal liver lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 18 kDa

Observed band size: 15 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-VKORC1 antibody [EPR20245] (AB206656)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-VKORC1 antibody [EPR20245] (ab206656) at 1/1000 dilution

Lane 1:

Mouse lung lysate at 10 µg

Lane 2:

Mouse pancreas lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 18 kDa

Observed band size: 15 kDa

false

Exposure time: 3min