Anti-FURIN antibody [RM2083]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-FURIN antibody [RM2083] (AB324440)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized K-562 (human chronic myelogenous leukemia lymphoblast) (Right) / Ramos (human Burkitt's lymphoma B lymphocyte) (Left) cells labelling FURIN with ab324440 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

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

Low expression : Ramos.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-FURIN antibody [RM2083] (AB324440)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized K-562 (human chronic myelogenous leukemia lymphoblast) cells labelling FURIN with ab324440 at 1/50 (10.0 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in K-562 and no staining in Ramos cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : Ramos.
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 (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 dilution.

• WB

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Western blot - Anti-FURIN antibody [RM2083] (AB324440)

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

Low expression : Ramos (PMID : 7931173).

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

Exposure time : Lane 1, 2 : 103 seconds Lane 3-6 : 37 seconds

All lanes:

Western blot - Anti-FURIN antibody [RM2083] (ab324440) at 1/1000 dilution

Lane 1:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lydate at 40 µg

Lane 2:

Ramos (human Burkitt's lymphoma B lymphocyte) whole cell lydate at 40 µg

Lane 3:

KARPAS-299 (human T cell lymphoma cell) whole cell lydate at 40 µg

Lane 4:

HEK-293 (human embryonic kidney epithelial cell) whole cell lydate at 40 µg

Lane 5:

Untreated KARPAS-299 whole cell lydate at 40 µg

Lane 6:

KARPAS-299 treated with Protein Deglycosylation MIX II 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: 85 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-FURIN antibody [RM2083] (AB324440)

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

Low expression : Human lung.

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

All lanes:

Western blot - Anti-FURIN antibody [RM2083] (ab324440) at 1/1000 dilution

Lane 1:

Human placenta tissue lysate at 20 µg

Lane 2:

Human lung 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: 85 kDa,36 kDa

false

Exposure time: 26s

• Dot

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Dot Blot - Anti-FURIN antibody [RM2083] (AB324440)

Dot blot analysis of FURIN using ab324440 at 1 : 1000 (0.5 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : No-tagged human PCSK3 recombinant protein
Lane2 : His-tagged human PCSK1 recombinant protein
Lane3 : His-tagged human PCSK4 recombinant protein
Lane4 : His-tagged human PCSK5 recombinant protein
Lane5 : His-tagged human PCSK6 recombinant protein

Exposure time : 180 seconds.

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

This antibody does not cross-react with human PCSK1, PCSK4, PCSK5, and PCSK6.

In Dot blot, anti-6X His tag® antibody [AD1.1.10] (ab15149) staining at 1/1000 dilution.

All lanes:

Dot Blot - Anti-FURIN antibody [RM2083] (ab324440) at 1/1000 dilution

Lane 1:

No-tagged human PCSK3 recombinant protein

Lane 2:

His-tagged human PCSK1 recombinant protein

Lane 3:

His-tagged human PCSK4 recombinant protein

Lane 4:

His-tagged human PCSK5 recombinant protein

Lane 5:

His-tagged human PCSK6 recombinant protein

Secondary

All lanes:

Dot 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

false

Exposure time: 180s