Anti-SMOC1 antibody [EPR26922-29]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SMOC1 antibody [EPR26922-29] (AB313569)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling SMOC1 with ab313569 at 1/100 (5.14 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 HeLa cell line. 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 1/200 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.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SMOC1 antibody [EPR26922-29] (AB313569)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling SMOC1 with ab313569 at 1/50 dilution (1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IP

Collaborator

Immunoprecipitation - Anti-SMOC1 antibody [EPR26922-29] (AB313569)

SMOC1 was immunoprecipitated from 0.35 mg Wild-type HeLa (human cervical adenocarcinoma epithelial cell) concentrated culture supernatant (ultrafiltration). with ab313569 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313569 at 1/1000 dilution. Lane 1 : Wild-type HeLa (human cervical adenocarcinoma epithelial cell) concentrated culture supernatant (ultrafiltration). Lane 2 : ab313569 IP in Wild-type HeLa (human cervical adenocarcinoma epithelial cell) concentrated culture supernatant (ultrafiltration). Lane 3 : Wild-type HeLa (human cervical adenocarcinoma epithelial cell) concentrated culture supernatant (ultrafiltration). Blocking and dilution buffer and concentration : 5% NFDM/TBST. The images were kindly provided by YCharOS Antibody Characterization.

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Immunoprecipitation - Anti-SMOC1 antibody [EPR26922-29] (ab313569) at 1/30 dilution

All lanes:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) concentrated culture supernatant (ultrafiltration)

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The images were kindly provided by YCharOS Antibody Characterization.

• IP

Supplier Data

Immunoprecipitation - Anti-SMOC1 antibody [EPR26922-29] (AB313569)

SMOC1 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) treated with 300 ng/ml Brefeldin A for 24 hours whole cell lysate with ab313569 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313569 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) treated with 300 ng/ml Brefeldin A for 24 hours whole cell lysate Lane 2 : ab313569 IP in HeLa (human cervical adenocarcinoma epithelial cell) treated with 300 ng/ml Brefeldin A for 24 hours whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313569 in HeLa treated with 300 ng/ml Brefeldin A for 24 hours whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 50 seconds

All lanes:

Immunoprecipitation - Anti-SMOC1 antibody [EPR26922-29] (ab313569) at 1/30 dilution

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HeLa (human cervical adenocarcinoma epithelial cell) treated with 300 ng/ml Brefeldin A for 24 hours whole cell lysate

Secondary

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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

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Exposure time: 50s

• WB

Collaborator

Western blot - Anti-SMOC1 antibody [EPR26922-29] (AB313569)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Cells were grown for 24 hours in serum free media before the media was collected. The images were kindly provided by YCharOS Antibody Characterization. Exposure time : 2 minutes

All lanes:

Western blot - Anti-SMOC1 antibody [EPR26922-29] (ab313569) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) cture supernatant (trafiltration) at 30 µg

Lane 2:

SMOC1 knockout HeLa cture supernatant (trafiltration) at 30 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/5000 dilution

Observed band size: 50-60 kDa

false

Exposure time: 2min

The images were kindly provided by YCharOS Antibody Characterization.

• WB

Supplier Data

Western blot - Anti-SMOC1 antibody [EPR26922-29] (AB313569)

Blocking and diluting buffer and concentration : 5% NFDM/TBST In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 48 seconds

All lanes:

Western blot - Anti-SMOC1 antibody [EPR26922-29] (ab313569) at 1/1000 dilution

Lane 1:

Untreated HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa treated with /ml Brefeldin A for 24 hours 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 1/20000 dilution

Observed band size: 50-60 kDa

false

Exposure time: 48s