Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Avian - Chicken, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, ELISA Capture (Matched Antibody Pair), Neutralization, Immunoprecipitation

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunoprecipitation, Luminex Development

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Cathepsin S Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Cathepsin S
Gln17-Ile331
Accession # P25774

Specificity

Detects human Cathepsin S in ELISAs and Western blots. In sandwich ELISAs, less than 0.05% cross-reactivity with recombinant human Cathepsin A, B, C, D, E, L, V, and X/Z/P is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human Cathepsin S Antibody

Cathepsin S antibody in Human Lymph Node by Immunohistochemistry (IHC-P).

Cathepsin S in Human Lymph Node.

Cathepsin S was detected in immersion fixed paraffin-embedded sections of human lymph node using Goat Anti-Human Cathepsin S Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1183) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of immersion fixed paraffin-embedded Tissue Sections.
Detection of Human Cathepsin S by Western Blot

Detection of Human Cathepsin S by Western Blot

CTSS attenuates EGF-mediated EGFR degradation.(a) OEC-M1 and MDA-MB-231 cells were pretreated with 20 μM 6r or ZFL for 1 h and subsequently incubated with 100 ng/mL EGF for an additional 2 h. The total cell lysates were analysed using EGFR-specific antibodies. ACTIN was used as the internal control for semiquantitative loading in each lane. (b) The cells were stimulated with EGF (100 ng/mL) with or without the pretreatment of 20 μM 6r for the indicated durations. EGFR degradation was examined through immunostaining by using an anti-EGFR antibody. Notably, a substantial amount of EGFR was detectable even after 6 h of EGF stimulation in 6r-treated cells. (c) The OEC-M1 cells were transiently transfected with plasmids (pCMV) that encoded wild-type CTSS. After 24 h of transfection, the cells were treated with 100 ng/mL EGF for the indicated durations and the cellular EGFR, CTSS, and ACTIN signals were determined through Western blotting. The lifespan of EGF-mediated EGFR degradation was calculated by normalising the signal intensity of EGFR with that of ACTIN. (d) The MDA-MB-231 cells were transfected with specific 50 nM CTSS siRNA (si-CTSS) for 24 h and subsequently incubated with 100 ng/mL EGF for the indicated durations. The nontargeting scramble siRNA (si-SC) was used as the scramble control. (e) The MDA-MB-231 cells were transiently transfected with plasmids encoding the CTSS-C25A mutant. After 24 h of transfection, the cells were incubated with 100 ng/mL of EGF for the indicated durations. Furthermore, the cells were harvested and subjected to SDS-PAGE and Western blotting. EGFR degradation was determined using an antibody against EGFR. ACTIN signalling was included as the loading control. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/srep29256), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Cathepsin S by Western Blot

Detection of Human Cathepsin S by Western Blot

CTSS inhibition does not impair lysosomal activities.(a) OEC-M1 cells were pretreated with vesicle or 100 nM BAF for 1 h and then incubated with 20 μM 6r for 1 h further. Lysosomal proteolytic activities were determined using BODIPY–BSA and quantified through flow cytometry. (b) After 48 h of siRNA knockdown of CTSS and CTSB, the relative expression of CTSS and CTSB were determined through Western blotting (right panel). Lysosomal proteolytic activities were determined using BODIPY–BSA and quantified through flow cytometry (right panel). Data represent the mean ± SD of three independent experiments. Differences were found to be statistically significant at ***P < 0.001. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/srep29256), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Cathepsin S by Immunocytochemistry/ Immunofluorescence

Detection of Mouse Cathepsin S by Immunocytochemistry/ Immunofluorescence

Myc induces cathepsin L expression in beta-cells of pancreatic Islets.(A) Immunohistochemical analyses for CTS B, C, L or S expression (all in red) in combination with staining for the pan-leukocyte marker CD45 (green) in pancreatic islet tumors from the MycERTAM;Bcl-xL animals. Pancreata were harvested from the MycERTAM;Bcl-xL mice treated for 7 d with TAM (Myc-On, 7 days) or control vehicle in place of TAM (Myc-OFF). The islet area is indicated by dotted line. The asterisks indicate the area of tumor represented in the insets. The panels are representatives of at least three animals assayed at each data point, all immunohistochemical analyses were done in duplicate; eight randomized fields per analysis were examined. Scale bars, 100μm. (B) Immunohistochemical analysis for cathepsin L expression in beta-cells of pancreatic islets from MycERTAM;Bcl-xL animals identified by insulin expression. Pancreata were collected from the animals described above. Scale bars represent 25μm. The panels are representatives of three animals assayed at each data point, all immunohistochemical analyses were done in duplicate; ten randomized fields per analysis were examined. Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0120348), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Cathepsin S by Immunocytochemistry/ Immunofluorescence

Detection of Mouse Cathepsin S by Immunocytochemistry/ Immunofluorescence

Myc induces cathepsin L expression in beta-cells of pancreatic Islets.(A) Immunohistochemical analyses for CTS B, C, L or S expression (all in red) in combination with staining for the pan-leukocyte marker CD45 (green) in pancreatic islet tumors from the MycERTAM;Bcl-xL animals. Pancreata were harvested from the MycERTAM;Bcl-xL mice treated for 7 d with TAM (Myc-On, 7 days) or control vehicle in place of TAM (Myc-OFF). The islet area is indicated by dotted line. The asterisks indicate the area of tumor represented in the insets. The panels are representatives of at least three animals assayed at each data point, all immunohistochemical analyses were done in duplicate; eight randomized fields per analysis were examined. Scale bars, 100μm. (B) Immunohistochemical analysis for cathepsin L expression in beta-cells of pancreatic islets from MycERTAM;Bcl-xL animals identified by insulin expression. Pancreata were collected from the animals described above. Scale bars represent 25μm. The panels are representatives of three animals assayed at each data point, all immunohistochemical analyses were done in duplicate; ten randomized fields per analysis were examined. Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0120348), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Cathepsin S by Western Blot

Detection of Cathepsin S by Western Blot

Proteolytic cleavage of EGFR by CTSS.(a) Recombinant EGFR was incubated with CTSS (left panel) and CTSB (right panel) at 37 °C for the indicated durations. The reaction was stopped by adding a sample buffer, and the reaction mixtures were subjected to SDS-PAGE, followed by Western blotting with an EGFR antibody. (b) Recombinant EGFR was incubated with CTSS in the presence of a CTSS inhibitor 6r or ZFL. The reaction mixtures were incubated at 37 °C for 20 min and then subjected to SDS-PAGE. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/27387133), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Cathepsin S by Western Blot

Detection of Cathepsin S by Western Blot

CTSS attenuates EGF-mediated EGFR degradation.(a) OEC-M1 and MDA-MB-231 cells were pretreated with 20 μM 6r or ZFL for 1 h and subsequently incubated with 100 ng/mL EGF for an additional 2 h. The total cell lysates were analysed using EGFR-specific antibodies. ACTIN was used as the internal control for semiquantitative loading in each lane. (b) The cells were stimulated with EGF (100 ng/mL) with or without the pretreatment of 20 μM 6r for the indicated durations. EGFR degradation was examined through immunostaining by using an anti-EGFR antibody. Notably, a substantial amount of EGFR was detectable even after 6 h of EGF stimulation in 6r-treated cells. (c) The OEC-M1 cells were transiently transfected with plasmids (pCMV) that encoded wild-type CTSS. After 24 h of transfection, the cells were treated with 100 ng/mL EGF for the indicated durations and the cellular EGFR, CTSS, and ACTIN signals were determined through Western blotting. The lifespan of EGF-mediated EGFR degradation was calculated by normalising the signal intensity of EGFR with that of ACTIN. (d) The MDA-MB-231 cells were transfected with specific 50 nM CTSS siRNA (si-CTSS) for 24 h and subsequently incubated with 100 ng/mL EGF for the indicated durations. The nontargeting scramble siRNA (si-SC) was used as the scramble control. (e) The MDA-MB-231 cells were transiently transfected with plasmids encoding the CTSS-C25A mutant. After 24 h of transfection, the cells were incubated with 100 ng/mL of EGF for the indicated durations. Furthermore, the cells were harvested and subjected to SDS-PAGE and Western blotting. EGFR degradation was determined using an antibody against EGFR. ACTIN signalling was included as the loading control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/27387133), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Cathepsin S Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human lymph node

Immunoprecipitation

25 µg/mL
Sample: Conditioned cell culture medium spiked with Recombinant Human Cathepsin S (Catalog # 1183-CY), see our available Western blot detection antibodies

Western Blot

0.1 µg/mL
Sample: Recombinant Human Cathepsin S (Catalog # 1183-CY)

Neutralization

Measured by its ability to neutralize activation and the resulting activity of Recombinant Human Cathepsin S (0.25 µg/mL, Catalog # 1183-CY) in cleaving the fluorogenic peptide substrate Mca-RPKPVE-Nval-WRK(Dnp)-NH2 (10 µM, Catalog # ES002). The Neutralization Dose (ND50) is typically 0.2-1.8 µg/mL.

Human Cathepsin S Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 0.2-0.8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Human Cathepsin S Biotinylated Antibody (Catalog # BAF1183)
  • Standard: Recombinant Human Cathepsin S Protein, CF (Catalog # 1183-CY)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 5 using AF1183 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Cathepsin S

Cathepsin S is a lysosomal cysteine protease of the papain family (1). It plays a major role in the processing of the MHC class II-associated invariant chain (2). It has been implicated in the pathogenesis of several diseases such as Alzheimer’s disease and degenerative disorders associated with the cells of the mononuclear phagocytic system (1). Human Cathepsin S is synthesized as a preproenzyme of 331 amino acid residues consisting a signal peptide (residues 1-16), a pro region (residues 17-114), and the mature enzyme (residues 115-331) (3-5). Cathepsin S is less abundant in tissues than Cathepsins B, L and H. The highest levels have been found in lymph nodes, spleen, macrophages, and other phagocytic cells.

References

  1. Kirschke, H. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, A.J. et al.) pp. 1104 - 1107, Academic Press, San Diego.
  2. Turk, V. et al. (2001) EMBO J. 20:4629.
  3. Shi, G.P. et al. (1992) J. Biol. Chem. 267:7258.
  4. Shi, G.P. et al. (1994) J. Biol. Chem. 269:11530.
  5. Wiederanders, B. et al. (1992) J. Biol. Chem. 267:13708.

Alternate Names

CTSS

Entrez Gene IDs

1520 (Human)

Gene Symbol

CTSS

UniProt

P25774

Additional Cathepsin S Products

Product Documents for Human Cathepsin S Antibody

Certificate of Analysis

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Product Specific Notices for Human Cathepsin S Antibody

For research use only

Citations for Human Cathepsin S Antibody

Customer Reviews for Human Cathepsin S Antibody (2)

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  • Human Cathepsin S Antibody
    Name: Anonymous
    Application: Simple Western
    Sample Tested: ARP-1 human myeloma cell line and JJN3 human myeloma cell line
    Species: Human
    Verified Customer | Posted 11/12/2020
    Human Cathepsin S Antibody AF1183
  • Human Cathepsin S Antibody
    Name: Balaji Mahender
    Application: ELISA
    Sample Tested: EDTA Plasma
    Species: Human
    Verified Customer | Posted 12/01/2017

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