Anti-Gli3 antibody [EPR26837-58]

• IP

Supplier Data

Immunoprecipitation - Anti-Gli3 antibody [EPR26837-58] (AB307714)

Gli3 was immunoprecipitated from 0.35 mg NIH/3T3 whole cell lysate with ab307714 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307714 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate Lane 2 : ab307714 IP in NIH/3T3 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307714 in NIH/3T3 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 32 seconds The lysate was freshly made and used for IP immediately to minimize protein degradation.

All lanes:

Immunoprecipitation - Anti-Gli3 antibody [EPR26837-58] (ab307714) at 1/30 dilution

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: 32s

• WB

Supplier Data

Western blot - Anti-Gli3 antibody [EPR26837-58] (AB307714)

Blocking and diluting buffer and concentration : 5% NFDM/TBST This antibody can detect the GLI3 full-length protein (190 kDa) and GLI3 C-terminally truncated form (83 kDa), which is consistent with that described in the literature (PMID : 20360384; PMID : 31554934). Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. Exposure time : 180 seconds

All lanes:

Western blot - Anti-Gli3 antibody [EPR26837-58] (ab307714) at 1/1000 dilution

Lane 1:

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

Lane 2:

293T (human embryonic kidney 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: 83 kDa,190 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Gli3 antibody [EPR26837-58] (AB307714)

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

This antibody can detect the GLI3 full-length protein (190 kDa) and GLI3 C-terminally truncated form (83 kDa), which is consistent with that described in the literature (PMID : 20360384; PMID : 31554934).

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

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.

In Western blot, anti- Vinculin antibody (ab129002) loading control staining at 1/10000 dilution.

All lanes:

Western blot - Anti-Gli3 antibody [EPR26837-58] (ab307714) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) transfected with scrambled siRNA control whole cell lysate at 50 µg

Lane 2:

NIH/3T3 transfected with siRNA specifically targeti GLI3 whole cell lysate at 50 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: 83 kDa,190 kDa

true

Exposure time: 48s