Anti-STIL/SIL antibody [EPR26955-72] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-STIL/SIL antibody [EPR26955-72] - BSA and Azide free (AB315886)

This data was developed using ab315885, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling STIL/SIL with ab315885 at 1/50 dilution (1 ug)/Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

HeLa cells are co-stained with DRAQ5 (PMID : 29352115).

• IP

Supplier Data

Immunoprecipitation - Anti-STIL/SIL antibody [EPR26955-72] - BSA and Azide free (AB315886)

This data was developed using ab315885, the same antibody clone in a different buffer formulation.

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

STIL/SIL was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab315885 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315885 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/1000 dilution.

Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab315885 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab315885 in HeLa whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-STIL/SIL antibody [EPR26955-72] (<a href='/en-us/products/primary-antibodies/stil-sil-antibody-epr26955-72-ab315885'>ab315885</a>) at 1/30 dilution

All lanes:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 10 µg

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 1/1000 dilution

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-STIL/SIL antibody [EPR26955-72] - BSA and Azide free (AB315886)

This data was developed using ab315885, the same antibody clone in a different buffer formulation.

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

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

All lanes:

Western blot - Anti-STIL/SIL antibody [EPR26955-72] (<a href='/en-us/products/primary-antibodies/stil-sil-antibody-epr26955-72-ab315885'>ab315885</a>) at 1/1000 dilution

Lanes 1 and 3:

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 1/20000 dilution

Observed band size: 175 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-STIL/SIL antibody [EPR26955-72] - BSA and Azide free (AB315886)

This data was developed using ab315885, the same antibody clone in a different buffer formulation.

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

Exposure time : Lanes 1-2 : 103 seconds; Lane 3 : 37 seconds.

All lanes:

Western blot - Anti-STIL/SIL antibody [EPR26955-72] (<a href='/en-us/products/primary-antibodies/stil-sil-antibody-epr26955-72-ab315885'>ab315885</a>) at 1/1000 dilution

Lane 1:

DU 145 (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Daudi (human burkitts lymphoma lymphoblast) 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: 175 kDa

true

• WB

Supplier Data

Western blot - Anti-STIL/SIL antibody [EPR26955-72] - BSA and Azide free (AB315886)

This data was developed using ab315885, 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.

All lanes:

Western blot - Anti-STIL/SIL antibody [EPR26955-72] (<a href='/en-us/products/primary-antibodies/stil-sil-antibody-epr26955-72-ab315885'>ab315885</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting STIL/SIL whole cell lysate at 20 µg

Lane 3:

K-562 (human chronic myelogenous leukemia lymphoblast) 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: 175 kDa,36 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-STIL/SIL antibody [EPR26955-72] - BSA and Azide free (AB315886)

This data was developed using ab315885, 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-STIL/SIL antibody [EPR26955-72] (<a href='/en-us/products/primary-antibodies/stil-sil-antibody-epr26955-72-ab315885'>ab315885</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 5 µg

Lane 2:

293T cells transfected with a human STIL/SIL expression vector containing a myc-His-tag®, whole cell lysate at 5 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: 175 kDa,36 kDa

false

Exposure time: 8s