Anti-Cathepsin S antibody [EPR5128]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin S antibody [EPR5128] (AB134157)

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] (AB134157)

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] (AB134157)

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] (ab134157) at 1/1000 dilution

Lanes 3 - 4:

Western blot - Anti-Cathepsin S antibody [EPR5128] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/cathepsin-s-antibody-epr5128-bsa-and-azide-free-ab248706'>ab248706</a>) 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] (AB134157)

All lanes:

Western blot - Anti-Cathepsin S antibody [EPR5128] (ab134157) 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

• WB

CiteAb

Western blot - Anti-Cathepsin S antibody [EPR5128] (AB134157)

Cathepsin S western blot using anti-Cathepsin S antibody [EPR5128] ab134157. Publication image and figure legend from Pawar, K., Sharbati, J., et al., 2016, Front Cell Infect Microbiol, PubMed 27014637.

ab134157 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab134157 please see the product overview.

Detection of autophagy in THP-1 cells transfected with miR-3619-5p mimics. (A) Expression of CTSS in THP-1 cells was down-regulated after transfection with miR-3619-5p mimics as well as siRNA, which affected the process of autophagy observed by LC3A/B conversion on western blots. (B) LC3 autophagy puncta were observed under the microscope, (C) and were significantly increased in miR-3619-5p mimics transfected cells compared to nonsense controls as calculated by ImageJ. (D) LC3-GFP autophagy puncta were also observed with pEX-GFP-hLC3-WT plasmids. Co-transfecting cells with miR-3619-5p with pEX-GFP-hLC3-WT plasmid produced more autophagy GFP-LC3 puncta than that of co-transfected cells with nonsense control and pEX-GFP-hLC3-WT plasmid as observed under fluorescent microscope. Arrow indicates co-transfected cells and arrowhead indicates non-transfected population. (E) Significant differences in number of autophagic GFP-LC3 puncta was observed between two groups counted by ImageJ. The columns show means of three biological replicates each measured in triplicates while error bars show the standard deviation. Asterisks indicate statistical significance between samples (*p < 0.05).

false