Anti-Cathepsin S antibody [EPR5128] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin S antibody [EPR5128] - BSA and Azide free (AB248706)

This data was developed using ab134157, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling Cathepsin S with purified ab134157 at 1/500 dilution (0.452 μg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin S antibody [EPR5128] - BSA and Azide free (AB248706)

This data was developed using ab134157, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue sections labeling Cathepsin S with purified ab134157 at 1/500 dilution (0.452 μg/mL). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• WB

Lab

Western blot - Anti-Cathepsin S antibody [EPR5128] - BSA and Azide free (AB248706)

False colour image of Western blot : Anti-Cathepsin S antibody [EPR5128] 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, ab134157 was shown to bind specifically to Cathepsin S. A band was observed at 25 kDa in wild-type THP-1 cell lysates with no signal observed at this size in Ctss knockout cell line ab275846 (knockout cell lysate ab275820). To generate this image, wild-type and Ctss knockout THP-1 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.

Lanes 1 - 2:

Western blot - Anti-Cathepsin S antibody [EPR5128] (<a href='/en-us/products/primary-antibodies/cathepsin-s-antibody-epr5128-ab134157'>ab134157</a>) at 1/1000 dilution

Lanes 3 - 4:

Western blot - Anti-Cathepsin S antibody [EPR5128] - BSA and Azide free (ab248706) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

Ctss knockout THP-1 cell lysate at 20 µg

Lane 2:

Western blot - Human CTSS knockout THP-1 cell line (<a href='/en-us/products/cell-lines/human-ctss-knockout-thp-1-cell-line-ab275846'>ab275846</a>)

Lane 3:

U-87 MG cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Predicted band size: 37 kDa

Observed band size: 25 kDa

false

• WB

Unknown

Western blot - Anti-Cathepsin S antibody [EPR5128] - BSA and Azide free (AB248706)

All lanes:

Western blot - Anti-Cathepsin S antibody [EPR5128] (<a href='/en-us/products/primary-antibodies/cathepsin-s-antibody-epr5128-ab134157'>ab134157</a>) at 1/1000 dilution

All lanes:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate

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

Predicted band size: 37 kDa

false