Anti-Cathepsin S antibody [EPR5128]

5 product images

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  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] (Anti-alpha Tubulin antibody [DM1A] - Loading Control 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 Human CTSS knockout THP-1 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed 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 (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 (Human CTSS knockout THP-1 cell line ab275846)

  Lane 3: U-87 MG cell lysate at 20 µg

  Lane 4: HeLa cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 37 kDa

  Observed band size: 25 kDa

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  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) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Predicted band size: 37 kDa

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  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 (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.
  The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

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  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 (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.
  The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

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  Image collected and cropped by CiteAb under a CC-BY license from the publication

  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).