Anti-Cripto1/CRIPTO antibody [RM1108] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Cripto1/CRIPTO antibody [RM1108] - BSA and Azide free (AB315249)

This data was developed using ab315248, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NCCIT (human pluripotent embryonic carcinoma epithelial cell) cells labelling Cripto1/CRIPTO with ab315248 at 1/50 (10.06 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 staining in NCCIT cell line. Negative control : MCF7.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/500 (1 ug/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-Cripto1/CRIPTO antibody [RM1108] - BSA and Azide free (AB315249)

This data was developed using ab315248, the same antibody clone in a different buffer formulation.

Cripto1/CRIPTO was immunoprecipitated from 0.35 mg NCCIT (human pluripotent embryonic carcinoma epithelial cell) whole cell lysate with ab315248 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315248 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : NCCIT (human pluripotent embryonic carcinoma epithelial cell) whole cell lysate

Lane 2 : ab315248 IP in NCCIT (human pluripotent embryonic carcinoma epithelial cell) whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab315248 in NCCIT whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-Cripto1/CRIPTO antibody [RM1108] (<a href='/en-us/products/primary-antibodies/cripto1-cripto-antibody-rm1108-ab315248'>ab315248</a>) at 1/30 dilution

All lanes:

NCCIT (human pluripotent embryonic 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: 15s

• WB

Supplier Data

Western blot - Anti-Cripto1/CRIPTO antibody [RM1108] - BSA and Azide free (AB315249)

This data was developed using ab315248, the same antibody clone in a different buffer formulation.

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

Low expression : MCF7(PMID : 25889638).

The double bands observed are consistent with what has been described in the literature. The lower band represents the immature form of Cripto1/CRIPTO. The upper band is likely to be the processed glycosylated form (PMID : 27881714 and 20568225).

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

All lanes:

Western blot - Anti-Cripto1/CRIPTO antibody [RM1108] (<a href='/en-us/products/primary-antibodies/cripto1-cripto-antibody-rm1108-ab315248'>ab315248</a>) at 1/1000 dilution

Lane 1:

NCCIT (human pluripotent embryonic carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) 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: 21 kDa,36 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-Cripto1/CRIPTO antibody [RM1108] - BSA and Azide free (AB315249)

This data was developed using ab315248, the same antibody clone in a different buffer formulation.

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

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

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-Cripto1/CRIPTO antibody [RM1108] (<a href='/en-us/products/primary-antibodies/cripto1-cripto-antibody-rm1108-ab315248'>ab315248</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a myc-His tag, whole cell lysate at 10 µg

Lane 2:

293T cells transfected with a human CRIPTO expression vector containing a His-tag, whole cell lysate at 10 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: 21 kDa,36 kDa

false

Exposure time: 8s