Anti-NOTUM antibody [EPR27981-76] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-NOTUM antibody [EPR27981-76] - BSA and Azide free (AB315896)

This data was developed using ab315895, the same antibody clone in a different buffer formulation.

NOTUM was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab315895 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315895 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 2 : ab315895 IP in HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab315895 in HepG2 whole cell lysate

All lanes:

Immunoprecipitation - Anti-NOTUM antibody [EPR27981-76] (<a href='/en-us/products/primary-antibodies/notum-antibody-epr27981-76-ab315895'>ab315895</a>) at 1/30 dilution

All lanes:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-NOTUM antibody [EPR27981-76] - BSA and Azide free (AB315896)

This data was developed using ab315895, the same antibody clone in a different buffer formulation.

The identity of the bands at 15 kDa and 70 kDa are unknown.

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

Exposure time : Lanes 1-2 : 180 seconds, lanes 3-4 : 39 seconds

All lanes:

Western blot - Anti-NOTUM antibody [EPR27981-76] (<a href='/en-us/products/primary-antibodies/notum-antibody-epr27981-76-ab315895'>ab315895</a>) at 1/1000 dilution

Lane 1:

Untreated SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

SW480 treated with 300ng/ml BFA for 24 hours whole cell lysate at 20 µg

Lane 3:

Untreated SW620 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

SW620 treated with 300ng/ml BFA for 24 hours 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 1/20000 dilution

Observed band size: 55 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-NOTUM antibody [EPR27981-76] - BSA and Azide free (AB315896)

This data was developed using ab315895, the same antibody clone in a different buffer formulation.

Low expression : Huh7 (PMID : 18429952).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 18429952).

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

All lanes:

Western blot - Anti-NOTUM antibody [EPR27981-76] (<a href='/en-us/products/primary-antibodies/notum-antibody-epr27981-76-ab315895'>ab315895</a>) at 1/1000 dilution

Lane 1:

Untreated HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HepG2 treated with 300ng/ml BFA for 24 hours whole cell lysate at 20 µg

Lane 3:

Untreated Huh7 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Huh7 treated with 300ng/ml BFA for 24 hours 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

Observed band size: 55 kDa,36 kDa

false

Exposure time: 92s

• WB

Supplier Data

Western blot - Anti-NOTUM antibody [EPR27981-76] - BSA and Azide free (AB315896)

This data was developed using ab315895, the same antibody clone in a different buffer formulation.

Low expression : lung (PMID : 18429952).

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

All lanes:

Western blot - Anti-NOTUM antibody [EPR27981-76] (<a href='/en-us/products/primary-antibodies/notum-antibody-epr27981-76-ab315895'>ab315895</a>) at 1/1000 dilution

Lane 1:

Human placenta tissue lysate at 20 µg

Lane 2:

Human lung tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 55 kDa,36 kDa

false

Exposure time: 180s