Anti-TSG6 antibody [EPR17582-45]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-TSG6 antibody [EPR17582-45] (AB267469)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized THP-1 (Human monocytic leukemia monocyte) (treated with 100ng/ml lipopolysaccharide (LPS) for 1h, then add BFA for 4h) (Red) / Untreated control (Green) cells labelling TSG6 with ab267469 at 1/500 dilution (0.1ug) (Red, Green) 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, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IP

Unknown

Immunoprecipitation - Anti-TSG6 antibody [EPR17582-45] (AB267469)

TSG6 was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte) (treated with 100ng/mg LPS for 3 hours, then treated with BFA for 2 hours) whole cell lysate with ab267469 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab267469 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : THP-1 (treated with 100ng/mg LPS for 3 hours, then treated with BFA for 2 hours) whole cell lysate 10 ug.

Lane 2 : ab267469 IP in THP-1 (treated as above).

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab267469 in THP-1 (treated as above) whole cell lysate

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

Exposure time : 41 seconds

This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Immunoprecipitation - Anti-TSG6 antibody [EPR17582-45] (ab267469)

Predicted band size: 31 kDa

Observed band size: 36 kDa

false

• WB

Lab

Western blot - Anti-TSG6 antibody [EPR17582-45] (AB267469)

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

Band of 36KDa represents the monomeric TSG6. Band of 120KDa likely represents a stable covalent complex formed between TSG6 and one of the heavy chains of inter-α-inhibitor or pre-α-inhibitor (PMID : 22351758, 11420253).

The expression profile observed is consistent with what has been described in the literature (PMID : 22351758, 11420253, 19389798).

Negative control : A549 (PMID : 19389798).

Exposure time : 3 minutes

All lanes:

Western blot - Anti-TSG6 antibody [EPR17582-45] (ab267469) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte), whole cell lysate at 20 µg

Lane 2:

THP-1 (treated with 100ng/mg LPS for 3 hours, then treated with BFA for 2 hours), whole cell lysate at 20 µg

Lane 3:

A549 (human lung carcinoma 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

Predicted band size: 31 kDa

Observed band size: 120 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-TSG6 antibody [EPR17582-45] (AB267469)

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

Band of 36KDa represents the monomeric TSG6. Band of 120KDa likely represents a stable covalent complex formed between TSG6 and one of the heavy chains of inter-α-inhibitor or pre-α-inhibitor (PMID : 22351758, 11420253).

The expression profile observed is consistent with what has been described in the literature (PMID : 22351758, 11420253, 19389798).

Exposure time : 3 minutes

All lanes:

Western blot - Anti-TSG6 antibody [EPR17582-45] (ab267469) at 1/1000 dilution

All lanes:

Rat amniotic membrane tissue 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

Predicted band size: 31 kDa

Observed band size: 120 kDa,36 kDa

false