Anti-Cleaved Notch1 (Val1744) antibody [EPR26110-518] - BSA and Azide free

• WB

Lab

Western blot - Anti-Cleaved Notch1 (Val1744) antibody [EPR26110-518] - BSA and Azide free (AB327387)

This data was developed using ab327383, the same antibody clone in a different buffer formulation.

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

Negative control : MCF7

To minimize protein degradation, cells were lysed immediately after harvest and lysates applied to a gel and transfer membrane for Western blotting as soon as possible.The identity of the lower MW bands are likely to be degradation fragments.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Cleaved Notch1 (Val1744) antibody [EPR26110-518] (ab327383) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 40 µg

Lane 2:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate at 40 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell 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: 110 kDa,36 kDa,300 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Cleaved Notch1 (Val1744) antibody [EPR26110-518] - BSA and Azide free (AB327387)

This data was developed using ab327383, the same antibody clone in a different buffer formulation.

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

DAPT is a γ-secretase inhibitor that blocks the cleavage and subsequent release of the Notch1 intracellular domain (NICD).

To minimize protein degradation, cells were lysed immediately after harvest and lysates applied to a gel and transfer membrane for Western blotting as soon as possible.The identity of the lower MW bands are likely to be degradation fragments.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Cleaved Notch1 (Val1744) antibody [EPR26110-518] (ab327383) at 1/1000 dilution

Lane 1:

Untreated Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 40 µg

Lane 2:

Jurkat treated with 0.1 μM DAPT for 24 hours whole cell lysate at 40 µg

Lane 3:

Jurkat treated with 1 μM DAPT for 24 hours whole cell 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: 110 kDa,36 kDa,300 kDa

true

Exposure time: 180s

• Dot

Lab

Dot Blot - Anti-Cleaved Notch1 (Val1744) antibody [EPR26110-518] - BSA and Azide free (AB327387)

This data was developed using ab327383, the same antibody clone in a different buffer formulation.

Dot blot analysis of Notch1 using ab327383 at 1 : 1000 (0.498 μg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane 1 : Cleaved Notch1 peptide (Val1744 neo-epitope)

Lane 2 : Non-cleaved control peptide (N-terminally extended)

Exposure time : 180 seconds.

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

The antibody specifically recognizes the cleaved peptide but not the non-cleaved form.

All lanes:

Dot Blot - Anti-Cleaved Notch1 (Val1744) antibody [EPR26110-518] (ab327383) at 1/1000 dilution

Lane 1:

Cleaved Notch1 peptide (Val1744 neo-epitope)

Lane 2:

Non-cleaved control peptide (N-terminally extended)

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

• WB

Lab

Western blot - Anti-Cleaved Notch1 (Val1744) antibody [EPR26110-518] - BSA and Azide free (AB327387)

This data was developed using ab327383, the same antibody clone in a different buffer formulation.

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

Low expression : EL4, Neuro-2a.

To minimize protein degradation, cells were lysed immediately after harvest and lysates applied to a gel and transfer membrane for Western blotting as soon as possible.The identity of the lower MW bands are likely to be degradation fragments.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Cleaved Notch1 (Val1744) antibody [EPR26110-518] (ab327383) at 1/1000 dilution

Lane 1:

HC11 (mouse mammary gland epithelial cell) whole cell lysate at 40 µg

Lane 2:

bEnd.3 (mouse brain endothelial cell) whole cell lysate at 40 µg

Lane 3:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 40 µg

Lane 4:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell 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: 110 kDa,36 kDa,300 kDa

false

Exposure time: 180s