Anti-DLL1 antibody [EPR29655-26]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-DLL1 antibody [EPR29655-26] (AB327088)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T cells transfected with an empty expression vector containing a myc-His-tag® HEK-293T cells transfected with a human Dll1 expression vector containing a myc-His-tag® HEK-293T cells transfected with a human Dll3 expression vector containing a myc-His-tag® HEK-293T cells transfected with a human Dll4 expression vector containing a myc-His-tag® cells labelling DLL1 with ab327088 at 1/1000 (0.509 μg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing positive staining in HEK-293T cells transfected with a human Dll1 expression vector containing a myc-His-tag®(shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). A : HEK-293T cells transfected with an empty expression vector containing a myc-His-tag®. B : HEK-293T cells transfected with a human Dll1 expression vector containing a myc-His-tag®. C : HEK-293T cells transfected with a human Dll3 expression vector containing a myc-His-tag®. D : HEK-293T cells transfected with a human Dll4 expression vector containing a myc-His-tag®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-DLL1 antibody [EPR29655-26] (AB327088)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized TT (human thyroid carcinoma epithelial cell) SH-SY5Y (human neuroblastoma epithelial cell) cells labelling DLL1 with ab327088 at 1/50 (10.18 μg/ml) dilution , followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing membranous and cytoplasmic staining in TT cells and no staining in SH-SY5Y cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Low expression : SH-SY5Y (PMID : 28525978). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab327088 at 1/50 dilution, followed by ab150120 at 1/1000 dilution.-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 at 1/1000 dilution.

• WB

Lab

Western blot - Anti-DLL1 antibody [EPR29655-26] (AB327088)

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

Low expression : SH-SY5Y (PMID : 28525978), SW620, AsPC-1.

The molecular weight observed is consistent with what has been described in the literature (PMID : 12794186).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-DLL1 antibody [EPR29655-26] (ab327088) at 1/1000 dilution

Lane 1:

TT (human thyroid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

FaDu (human pharyngeal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

AsPC-1 (human Pancreas epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 85 kDa,30 kDa,124 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-DLL1 antibody [EPR29655-26] (AB327088)

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

This antibody does not cross-react with overexpressed human DLL3 or DLL4 by western blot.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(85, 30KDa); Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-DLL1 antibody [EPR29655-26] (ab327088) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human DLL1 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human DLL3 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Lane 4:

293T cells transfected with a human DLL4 expression vector containing a myc-His-tag® whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 85 kDa,30 kDa,36 kDa

false

Exposure time: 8s

• WB

Lab

Western blot - Anti-DLL1 antibody [EPR29655-26] (AB327088)

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

Low expression : NIH/3T3, EL4.

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

All lanes:

Western blot - Anti-DLL1 antibody [EPR29655-26] (ab327088) at 1/1000 dilution

Lane 1:

bEnd.3 (mouse brain endothelial cell) whole cell lysate at 20 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 3:

EL4 (mouse lymphoma t lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 85 kDa,30 kDa,36 kDa

false

Exposure time: 180s