Anti-Furin antibody

• ICC

AbReview39690****

Immunocytochemistry - Anti-Furin antibody (AB3467)

ab3467 staining Furin in human A549 cells by Immunocytochemistry/ Immunofluorescence.

Cells were fixed with paraformaldehyde, permeabilized using 0.1% Triton X-100, blocked with 1% BSA/ 5%FBS/PBS for 1 hour at 25°C and then incubated with ab3467 at a 1/100 dilution for 1 hour at 25°C. The secondary used was an Alexa-Fluor 546 conjugated goat anti-rabbit polyclonal used at a 1/500 dilution.

Cells were counterstained with DAPI (436nM, 5 mins) and Phalloidin-633 (1unit/coverslip) and mounted with prolong gold. No staining seen in secondary control and furin staining was punctate and throughout the cytoplasm.

Image courtesy of an anonymous Abreview.

• ICC

Supplier Data

Immunocytochemistry - Anti-Furin antibody (AB3467)

Immunocytochemical analysis of methanol-fixed 2% BSA blocked HeLa cells strining furin with ab3467 at 2 μg/ml and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate at 1/2000 dilution (green). Nuclei are stained with SlowFade® Gold Antifade Mountant with DAPI (blue), and the cytoskeleton with alpha-Tubulin Monoclonal Antibody and Goat anti-Mouse IgG Secondary Antibody, Alexa Fluor® 594 conjugate at 1/400 dilution (red). Negative control uses no primary antibody

• ICC

Supplier Data

Immunocytochemistry - Anti-Furin antibody (AB3467)

Immunolocalization of endogenous furin in mouse 3T3 cells with ab3467.

• WB

Lab

Western blot - Anti-Furin antibody (AB3467)

Western blot : Rabbit Polyclonal to Furin ab3467 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 87 kDa in Wild-type A549 cell lysates with no signal observed at this size in FURIN knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Furin antibody (ab3467) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

FURIN knockout A549 at 20 µg

Lane 3:

HEK-293 at 20 µg

Lane 4:

MOLT-4 at 20 µg

Lane 5:

Ramos at 20 µg

Secondary

All lanes:

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

Predicted band size: 87 kDa

Observed band size: 87 kDa

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• WB

Supplier Data

Western blot - Anti-Furin antibody (AB3467)

All lanes:

Western blot - Anti-Furin antibody (ab3467) at 1/500 dilution

Lane 1:

HeLa lysate at 30 µg

Lane 2:

HEK-293 extract at 30 µg

Lane 3:

K-562 extract at 30 µg

Lane 4:

SH-SY5Y extract at 30 µg

Lane 5:

conditioned media from HeLa cell line at 10 µg

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate at 1/2500 dilution

Predicted band size: 87 kDa

true

• WB

Unknown

Western blot - Anti-Furin antibody (AB3467)

The band observed at 63 kDa could potentially be a cleaved form of Furin due to the presence of a 24 amino acid signal peptide and a 83 amino acid propeptide.

All lanes:

Western blot - Anti-Furin antibody (ab3467) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 87 kDa

Observed band size: 63 kDa

true

Exposure time: 90s

• WB

AbReview73973****

Western blot - Anti-Furin antibody (AB3467)

4-20% tris-glycine, denaturing conditions

All lanes:

Western blot - Anti-Furin antibody (ab3467) at 1/1000 dilution

Lane 1:

HAP1 at 25000 Cells

Lanes 2 - 3:

independent furin CRISPR KO clones (Horizon) at 25000 Cells

Secondary

All lanes:

rabbit IgG HRP Linked Whole Ab at 1/10000 dilution

true

Exposure time: 1min

This image is courtesy of an Abreview submitted by Lawrence Welch