Anti-FXI antibody [EPR29204-87]

• WB

Supplier Data

Western blot - Anti-FXI antibody [EPR29204-87] (AB323450)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature. The antibody detects the pro (XI) and active form (XIa) of FXI. PMID : 23515926, PMID : 23515926.

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

All lanes:

Western blot - Anti-FXI antibody [EPR29204-87] (ab323450) at 1/1000 dilution

Lane 1:

Untreated HepG2 whole cell lysate at 80 µg

Lane 2:

HepG2 treated with 300ng/ml BFA for 18 hours whole cell lysate at 80 µg

Secondary

All lanes:

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

Observed band size: 30-37 kDa,70-75 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-FXI antibody [EPR29204-87] (AB323450)

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

Low expression : HeLa, MCF7.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 29223926).

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

All lanes:

Western blot - Anti-FXI antibody [EPR29204-87] (ab323450) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 80 µg

Lane 2:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 80 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 80 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: 30-37 kDa,70-75 kDa,36 kDa

false

Exposure time: 70s

• WB

Supplier Data

Western blot - Anti-FXI antibody [EPR29204-87] (AB323450)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 29223926).

The identity of the band higher 250 kDa (in lanes 1-2) is unknown.

All lanes:

Western blot - Anti-FXI antibody [EPR29204-87] (ab323450) at 1/1000 dilution

Lane 1:

Human plasma lysate at 20 µg

Lane 2:

Human serum lysate at 20 µg

Lane 3:

Human liver tissue lysate at 80 µg

Secondary

All lanes:

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

Observed band size: 30-37 kDa,70-75 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-FXI antibody [EPR29204-87] (AB323450)

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

FXI is a glycoprotein of approximately 30-37, 70-75 kDa and is detected at a slightly lower molecular weight following N-glycosidase F (PNGase F) treatment.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 29223926).

For deglycosylation experiments, cells were incubated at 100°C for 10 mins, followed by incubation at 37°C for 1hr with or without N-glycosidase F (PNGase F).

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

All lanes:

Western blot - Anti-FXI antibody [EPR29204-87] (ab323450) at 1/1000 dilution

Lane 1:

Untreated HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 80 µg

Lane 2:

HepG2 whole cell lysate treated with Peptide: N-glycosidase F (PNGase F) at 80 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: 30-37 kDa,70-75 kDa,36 kDa

false

Exposure time: 70s

• Dot

Supplier Data

Dot Blot - Anti-FXI antibody [EPR29204-87] (AB323450)

Dot blot analysis of FXI using ab323450 at 1000 (0.493 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : LIF-tagged Human FXI light chain fragment
Lane2 : LIF-tagged Human FXI heavy chain fragment
Lane3 : LIF-tagged Human KLKB1 fragment

Exposure time : 180 seconds.

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

This antibody does not cross-react with human FX1 heavy chain and Human KLKB1.

Total protein control : Anti-LIF antibody [EPR21713-25] - BSA and Azide free (Capture) (ab244795) (1 : 1000).

All lanes:

Dot Blot - Anti-FXI antibody [EPR29204-87] (ab323450) at 1/1000 dilution

Lane 1:

LIF-tagged Human FXI light chain fragment

Lane 2:

LIF-tagged Human FXI heavy chain fragment

Lane 3:

LIF-tagged Human KLKB1 fragment

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