Anti-GAS 6 antibody [EPR23931-27]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-GAS 6 antibody [EPR23931-27] (AB264098)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized THP-1 (Human monocytic leukemia monocyte) cells labelling GAS 6 with ab264098 at 1/500 dilution (Right) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Cells were stained with rabbit IgG or ab264098.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-GAS 6 antibody [EPR23931-27] (AB264098)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized THP-1 (Human monocytic leukemia monocyte) cells labelling GAS 6 with ab264098 at 1/500 dilution (0.1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Cells were stained with rabbit IgG or ab264098. Cells were co-stained with DRAQ/ Ki-67 to differentiate cell cycle phase.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-GAS 6 antibody [EPR23931-27] (AB264098)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized Jurkat (Human T cell leukemia T lymphocyte (Left) / THP-1 (Human monocytic leukemia monocyte, Right) cells labelling GAS 6 with ab264098 at 1/500 dilution (Right). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Negative control : Jurkat (PMID : 10400186)

• IP

Supplier Data

Immunoprecipitation - Anti-GAS 6 antibody [EPR23931-27] (AB264098)

GAS 6 was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte) whole cell lysate with ab264098 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab264098 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : THP-1 (human monocytic leukemia monocyte) whole cell lysate 10 ug

Lane 2 : ab264098 IP in THP-1 whole cell lysate

Lane 3 : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated : Rabbit monoclonal IgG (ab172730) instead of ab264098 in THP-1 whole cell lysate

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

Exposure time : 15 seconds

All lanes:

Immunoprecipitation - Anti-GAS 6 antibody [EPR23931-27] (ab264098)

Predicted band size: 80 kDa

Observed band size: 78 kDa

false

• WB

Lab

Western blot - Anti-GAS 6 antibody [EPR23931-27] (AB264098)

False colour image of Western blot : Anti-GAS 6 antibody [EPR23931-27] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab264098 was shown to bind specifically to GAS 6. A band was observed at 75 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in GAS6 knockout cell line ab275837 (knockout cell lysate ab275811). To generate this image, wild-type and GAS6 knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-GAS 6 antibody [EPR23931-27] (ab264098) at 1/1000 dilution

Lane 1:

Wild-type HEK-293 cell lysate at 20 µg

Lane 2:

GAS6 knockout HEK-293 cell lysate at 20 µg

Lane 2:

Western blot - Human GAS6 knockout HEK-293 cell line (<a href='/en-us/products/cell-lines/human-gas6-knockout-hek-293-cell-line-ab275837'>ab275837</a>)

Lane 3:

THP-1 cell lysate at 20 µg

Predicted band size: 80 kDa

Observed band size: 75 kDa

false

• WB

Lab

Western blot - Anti-GAS 6 antibody [EPR23931-27] (AB264098)

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

Negative control : Jurkat (PMID : 10400186)

Samples are non-boiled as boiling may cause protein aggregates.

Exposure time : Lanes 1-2 : 48 seconds;Lanes 3-4 : 3 minutes.

All lanes:

Western blot - Anti-GAS 6 antibody [EPR23931-27] (ab264098) at 1/1000 dilution

Lane 1:

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

Lane 2:

Jurkat (human t cell leukemia T lymphocyte) whole cell lysate at 20 µg

Lane 3:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

RPMI-8226 (human plasmacytoma, myeloma B lymphocyte) 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 1/50000 dilution

Predicted band size: 80 kDa

Observed band size: 78 kDa

false