Anti-Jagged1 antibody [EPR4290] - BSA and Azide free

• WB

Lab

Western blot - Anti-Jagged1 antibody [EPR4290] - BSA and Azide free (AB184785)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109536).

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

Negative controls : Jurkat and MOLT-4 whole cell lysate (PMID : 30231940)

200-kDa full length and 23-kDa C-terminal JAG1 are observed. The molecular weights are consistent with what has been described in the literature (PMID : 30890522, 30890522).

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-Jagged1 antibody [EPR4290] (<a href='/en-us/products/primary-antibodies/jagged1-antibody-epr4290-ab109536'>ab109536</a>) at 1/10000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate

Lane 2:

HUVEC (human umbilical vein endothelial cell), whole cell lysate

Lane 3:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate

Lane 4:

MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate

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

Predicted band size: 134 kDa

Observed band size: 23 kDa,200 kDa

true

Exposure time: 180s

• WB

Lab

Western blot - Anti-Jagged1 antibody [EPR4290] - BSA and Azide free (AB184785)

Lanes 1 - 2 : Merged signal (red and green). Green - ab109536 observed at 200 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab109536 was shown to recognize in wild-type HAP1 cells as signal was lost at the expected MW in JAG1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and JAG1 knockout samples were subjected to SDS-PAGE. ab109536 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109536).

All lanes:

Western blot - Anti-Jagged1 antibody [EPR4290] (<a href='/en-us/products/primary-antibodies/jagged1-antibody-epr4290-ab109536'>ab109536</a>) at 1/10000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

JAG1 knockout HAP1 whole cell lysate at 20 µg

Predicted band size: 134 kDa

Observed band size: 200 kDa

false