重组Anti-SEPP1抗体[EPR24174-24]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SEPP1 antibody [EPR24174-24] (AB277526)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling SEPP1 with ab277526 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SEPP1 antibody [EPR24174-24] (AB277526)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 cells labelling SEPP1 with ab277526 at 1/50 (12.3 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HepG2 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-SEPP1 antibody [EPR24174-24] (AB277526)

SEPP1 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate with ab277526 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab277526 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate 10 ug

Lane 2 : ab277526 IP in HepG2 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab277526 in HepG2 whole cell lysate

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

Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-SEPP1 antibody [EPR24174-24] (ab277526)

Predicted band size: 43 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-SEPP1 antibody [EPR24174-24] (AB277526)

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

The observed MW is consistent with what has been described in the literature (PMID : 19345254, 27859223).

Exposure time : 10 seconds.

All lanes:

Western blot - Anti-SEPP1 antibody [EPR24174-24] (ab277526) at 1/1000 dilution

All lanes:

Human plasma at 10 µL

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 43 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-SEPP1 antibody [EPR24174-24] (AB277526)

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

The observed MW is consistent with what has been described in the literature (PMID : 19345254, 27859223).

This blot was developed using a higher sensitivity ECL substrate.

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-SEPP1 antibody [EPR24174-24] (ab277526) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellar carcinoma epithelial cell) (treated with 300ng/ml Brefeldin A (BFA) for 24 hours) whole cell lysate at 15 µg

Lane 2:

HepG2 (treated with 300ng/ml Brefeldin A (BFA) for 24 hours) whole cell lysate treated with PNGase F at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 43 kDa

Observed band size: 35-47 kDa,50-60 kDa

false

• WB

Lab

Western blot - Anti-SEPP1 antibody [EPR24174-24] (AB277526)

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

The observed MW is consistent with what has been described in the literature (PMID : 19345254, 27859223).

Low expression : Human skeletal muscle(PMID : 23038251).

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-SEPP1 antibody [EPR24174-24] (ab277526) at 1/1000 dilution

Lane 1:

Human liver tissue lysate at 20 µg

Lane 2:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

VeriBlot for IP secondary antibody(HRP)(<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 43 kDa

Observed band size: 50 kDa

false