Anti-Jagged1 antibody [EPR26134-17]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Jagged1 antibody [EPR26134-17] (AB300561)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Hepa1-6 (mouse hepatoma epithelial cell) cells labelling Jagged1 with ab300561 at 1/500 (1.04 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic and membranous staining in Hepa1-6 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). 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 2 ug/ml dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-Jagged1 antibody [EPR26134-17] (AB300561)

Jagged1 was immunoprecipitated from 0.35 mg Hepa1-6 (mouse hepatoma epithelial cell), whole cell lysate 10 ug with ab300561 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab300561 at ID270 dilution.

VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

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).

All lanes:

Immunoprecipitation - Anti-Jagged1 antibody [EPR26134-17] (ab300561) at 1/1000 dilution

Lane 1:

Hepa1-6 whole cell lysate at 10 µg

Lane 2:

ab300561 at 1/30 IP in Hepa1-6 whole cell lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab300561 in Hepa1-6 whole cell lysate at 10 µg

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

Observed band size: 23 kDa,200 kDa

false

Exposure time: 7.75s

• WB

Supplier Data

Western blot - Anti-Jagged1 antibody [EPR26134-17] (AB300561)

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

Negative controls : Jurkat, MOLT-4 (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 [EPR26134-17] (ab300561) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellar carcinoma epithelial cell), whole cell lysate 20

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 1/100000 dilution

Observed band size: 23 kDa,200 kDa

true

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Jagged1 antibody [EPR26134-17] (AB300561)

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

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).

Lane 3 of 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 [EPR26134-17] (ab300561) at 1/1000 dilution

Lane 1:

Hepa1-6 (mouse hepatoma epithelial cell), whole cell lysate

Lane 2:

bEnd.3 (mouse brain endothelioma), whole cell lysate

Lane 3:

AR42J (rat pancreatic tumor epithelial cell), 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 1/100000 dilution

Observed band size: 200 kDa,23 kDa

true