Anti-TMPRSS2 antibody [P5H9-A3] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TMPRSS2 antibody [P5H9-A3] - BSA and Azide free (AB309547)

This data was developed using ab309546 the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized LNCaP (human prostate carcinoma epithelial cell) cells labelling TMPRSS2 with ab309546 at 1/100 (10.28 ug/ml) dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in LNCaP and negative staining in 293T (PMID : 33493182).Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue). antibody only control : Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-TMPRSS2 antibody [P5H9-A3] - BSA and Azide free (AB309547)

This data was developed using ab309546, the same antibody clone in a different buffer formulation.

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

Negative control : 293T (PMID : 33493182).

In Western blot, anti- GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

All lanes:

Western blot - Anti-TMPRSS2 antibody [P5H9-A3] (<a href='/ko/products/primary-antibodies/tmprss2-antibody-p5h9-a3-ab309546'>ab309546</a>) at 1/1000 dilution

Lane 1:

LNCaP (human prostate carcinoma 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:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 25 kDa,55 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-TMPRSS2 antibody [P5H9-A3] - BSA and Azide free (AB309547)

This data was developed using ab309546, the same antibody clone in a different buffer formulation.Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of TBS Lysates at 20 µg per lane. The samples were run on a Bis-Tris gel under reducing conditions. Western blot : Anti-TMPRSS2 antibody (ab309546) staining at 1/1000 dilution, shown in green; Rabbit anti-GAPDH antibody (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab309546 was shown to bind specifically to TMPRSS2. Target of interest was observed at 55 kDa and 25 KDa in wild-type LNCaP cell lysates (lane 1) with no signal observed at this size in TMPRSS2 knockout cell line (lane 2, knockout cell line ab273745). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged. The molecular weight observed is consistent with what has been described in the literature (PMID : 35104687). Exposure time :

All lanes:

Western blot - Anti-TMPRSS2 antibody [P5H9-A3] (<a href='/ko/products/primary-antibodies/tmprss2-antibody-p5h9-a3-ab309546'>ab309546</a>) at 1/1000 dilution

Lane 1:

Wild-type LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

TMPRSS2 knockout LNCaP whole cell lysate at 20 µg

Lane 2:

Western blot - Human TMPRSS2 knockout LNCaP cell line (<a href='/ko/products/cell-lines/human-tmprss2-knockout-lncap-cell-line-ab273745'>ab273745</a>)

Secondary

Lanes 1 - 2:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/200000 dilution

Lanes 1 - 2:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/200000 dilution

Observed band size: 25 kDa,55 kDa

false