Matriptase/ST14 Antibody [Unconjugated]

HAF016). Specific bands were detected for Matriptase/ST14 at approximately 25 and 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1." > HAF016). Specific bands were detected for Matriptase/ST14 at approximately 25 and 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1." title="Western Blot shows lysates of HCT‑116 human colorectal carcinoma cell line and Ramos human Burkitt's lymphoma cell line. PVDF membrane was probed with 1 µg/ml of Sheep Anti-Human Matriptase/ST14 Catalytic Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3946) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for Matriptase/ST14 at approximately 25 and 90 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1." />

Western Blot shows lysates of HCT‑116 human colorectal carcinoma cell line and Ramos human Burkitt's lymphoma cell line. PVDF membrane was probed with 1 µg/ml of Sheep Anti-Human Matriptase/ST14 Catalytic Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3946) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # <a class=

HAF016) in Milk-free Antibody Diluent (Catalog # 043-524). This experiment was conducted under reducing conditions and using the 12-230kDa separation system. Right: Simple Western electropherogram showing the same Sheep Anti-Human Matriptase/ST14 Catalytic Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3946) tested at 10 µg/ml (blue line) and 50 µg/ml (green line) in the Ramos human Burkitt's lymphoma cell line." > HAF016) in Milk-free Antibody Diluent (Catalog # 043-524). This experiment was conducted under reducing conditions and using the 12-230kDa separation system. Right: Simple Western electropherogram showing the same Sheep Anti-Human Matriptase/ST14 Catalytic Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3946) tested at 10 µg/ml (blue line) and 50 µg/ml (green line) in the Ramos human Burkitt's lymphoma cell line." title="Left: Simple Western lane view shows lysates of Ramos human Burkitt's lymphoma cell line, loaded at 0.1 mg/ml. A specific band was detected for Matriptase/ST14 at approximately 96 kDa (as indicated) using both 10 µg/ml and 50 µg/ml of Sheep Anti-Human Matriptase/ST14 Catalytic Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3946) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016) in Milk-free Antibody Diluent (Catalog # 043-524). This experiment was conducted under reducing conditions and using the 12-230kDa separation system. Right: Simple Western electropherogram showing the same Sheep Anti-Human Matriptase/ST14 Catalytic Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3946) tested at 10 µg/ml (blue line) and 50 µg/ml (green line) in the Ramos human Burkitt's lymphoma cell line." />

<b>Left:</b> Simple Western lane view shows lysates of Ramos human Burkitt's lymphoma cell line, loaded at 0.1 mg/ml. A specific band was detected for Matriptase/ST14 at approximately 96 kDa (as indicated) using both 10 µg/ml and 50 µg/ml of Sheep Anti-Human Matriptase/ST14 Catalytic Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3946) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # <a class=

5-001-A, open histogram), followed by NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (NL010). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin." > 5-001-A, open histogram), followed by NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (NL010). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin." title="PC-3 human prostate cancer cell line was stained with Sheep Anti-Human Matriptase/ST14 Catalytic Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3946, filled histogram) or control antibody (5-001-A, open histogram), followed by NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (NL010). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin." />

PC-3 human prostate cancer cell line was stained with Sheep Anti-Human Matriptase/ST14 Catalytic Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3946, filled histogram) or control antibody (<a class=

Zymogen-locked matriptase induces epidermal prostasin processing. Protein extracts from skin (a), kidney (b), lung (c), and intestine (d) from newborn St14zym/zym (lanes 1 and 2), St14+/+ (lanes 3 and 4), and St14–/– (lane 5) littermates were separated by capillary electrophoresis and probed with antibodies against matriptase (top panels), prostasin (middle panels), or beta -actin (bottom panels). Lanes 6 and 7 in (a) are skin extracts from prostasin null (Prss8–/–) and prostasin zymogen-locked (Prss8zym/zym) mice, respectively. Zymogens of matriptase and prostasin are indicated with filled arrows, and the activated forms are indicated with open arrows. n.s. non-specific. Positions of molecular weight markers (kDa) are indicated on the left. e Representative example of quantification of activated prostasin (open arrow) and zymogen prostasin (filled arrows) in protein extracts from skin from a newborn St14zym/zym mouse (top panel), St14+/+ mouse (second panel from top), and a newborn St14–/– mouse (second panel from bottom). Skin extracts from a newborn mouse expressing zymogen-locked (Prss8zym/zym) endogenous prostasin is included as reference (bottom panel). f Ratio of activated prostasin to total prostasin in skin extracts from newborn St14zym/zym (left bar, n = 7), St14+/+ (middle bar, n = 7), and St14–/– (right bar, n = 2) mice, quantified as in (e). Data are shown as mean ± SD. *P = 0.0011 was determined by one-way ANOVA, two-tailed. Additional file 1: Raw supporting data Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28571576), licensed under a CC-BY license. Not internally tested by R&D Systems.

Zymogen-locked matriptase induces epidermal prostasin processing. Protein extracts from skin (a), kidney (b), lung (c), and intestine (d) from newborn St14zym/zym (lanes 1 and 2), St14+/+ (lanes 3 and 4), and St14–/– (lane 5) littermates were separated by capillary electrophoresis and probed with antibodies against matriptase (top panels), prostasin (middle panels), or beta -actin (bottom panels). Lanes 6 and 7 in (a) are skin extracts from prostasin null (Prss8–/–) and prostasin zymogen-locked (Prss8zym/zym) mice, respectively. Zymogens of matriptase and prostasin are indicated with filled arrows, and the activated forms are indicated with open arrows. n.s. non-specific. Positions of molecular weight markers (kDa) are indicated on the left. e Representative example of quantification of activated prostasin (open arrow) and zymogen prostasin (filled arrows) in protein extracts from skin from a newborn St14zym/zym mouse (top panel), St14+/+ mouse (second panel from top), and a newborn St14–/– mouse (second panel from bottom). Skin extracts from a newborn mouse expressing zymogen-locked (Prss8zym/zym) endogenous prostasin is included as reference (bottom panel). f Ratio of activated prostasin to total prostasin in skin extracts from newborn St14zym/zym (left bar, n = 7), St14+/+ (middle bar, n = 7), and St14–/– (right bar, n = 2) mice, quantified as in (e). Data are shown as mean ± SD. *P = 0.0011 was determined by one-way ANOVA, two-tailed. Additional file 1: Raw supporting data Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28571576), licensed under a CC-BY license. Not internally tested by R&D Systems.

Zymogen-locked matriptase induces epidermal prostasin processing. Protein extracts from skin (a), kidney (b), lung (c), and intestine (d) from newborn St14zym/zym (lanes 1 and 2), St14+/+ (lanes 3 and 4), and St14–/– (lane 5) littermates were separated by capillary electrophoresis and probed with antibodies against matriptase (top panels), prostasin (middle panels), or beta -actin (bottom panels). Lanes 6 and 7 in (a) are skin extracts from prostasin null (Prss8–/–) and prostasin zymogen-locked (Prss8zym/zym) mice, respectively. Zymogens of matriptase and prostasin are indicated with filled arrows, and the activated forms are indicated with open arrows. n.s. non-specific. Positions of molecular weight markers (kDa) are indicated on the left. e Representative example of quantification of activated prostasin (open arrow) and zymogen prostasin (filled arrows) in protein extracts from skin from a newborn St14zym/zym mouse (top panel), St14+/+ mouse (second panel from top), and a newborn St14–/– mouse (second panel from bottom). Skin extracts from a newborn mouse expressing zymogen-locked (Prss8zym/zym) endogenous prostasin is included as reference (bottom panel). f Ratio of activated prostasin to total prostasin in skin extracts from newborn St14zym/zym (left bar, n = 7), St14+/+ (middle bar, n = 7), and St14–/– (right bar, n = 2) mice, quantified as in (e). Data are shown as mean ± SD. *P = 0.0011 was determined by one-way ANOVA, two-tailed. Additional file 1: Raw supporting data Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28571576), licensed under a CC-BY license. Not internally tested by R&D Systems.

Zymogen-locked matriptase induces epidermal prostasin processing. Protein extracts from skin (a), kidney (b), lung (c), and intestine (d) from newborn St14zym/zym (lanes 1 and 2), St14+/+ (lanes 3 and 4), and St14–/– (lane 5) littermates were separated by capillary electrophoresis and probed with antibodies against matriptase (top panels), prostasin (middle panels), or beta -actin (bottom panels). Lanes 6 and 7 in (a) are skin extracts from prostasin null (Prss8–/–) and prostasin zymogen-locked (Prss8zym/zym) mice, respectively. Zymogens of matriptase and prostasin are indicated with filled arrows, and the activated forms are indicated with open arrows. n.s. non-specific. Positions of molecular weight markers (kDa) are indicated on the left. e Representative example of quantification of activated prostasin (open arrow) and zymogen prostasin (filled arrows) in protein extracts from skin from a newborn St14zym/zym mouse (top panel), St14+/+ mouse (second panel from top), and a newborn St14–/– mouse (second panel from bottom). Skin extracts from a newborn mouse expressing zymogen-locked (Prss8zym/zym) endogenous prostasin is included as reference (bottom panel). f Ratio of activated prostasin to total prostasin in skin extracts from newborn St14zym/zym (left bar, n = 7), St14+/+ (middle bar, n = 7), and St14–/– (right bar, n = 2) mice, quantified as in (e). Data are shown as mean ± SD. *P = 0.0011 was determined by one-way ANOVA, two-tailed. Additional file 1: Raw supporting data Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28571576), licensed under a CC-BY license. Not internally tested by R&D Systems.

Zymogen-locked matriptase induces epidermal prostasin processing. Protein extracts from skin (a), kidney (b), lung (c), and intestine (d) from newborn St14zym/zym (lanes 1 and 2), St14+/+ (lanes 3 and 4), and St14–/– (lane 5) littermates were separated by capillary electrophoresis and probed with antibodies against matriptase (top panels), prostasin (middle panels), or beta -actin (bottom panels). Lanes 6 and 7 in (a) are skin extracts from prostasin null (Prss8–/–) and prostasin zymogen-locked (Prss8zym/zym) mice, respectively. Zymogens of matriptase and prostasin are indicated with filled arrows, and the activated forms are indicated with open arrows. n.s. non-specific. Positions of molecular weight markers (kDa) are indicated on the left. e Representative example of quantification of activated prostasin (open arrow) and zymogen prostasin (filled arrows) in protein extracts from skin from a newborn St14zym/zym mouse (top panel), St14+/+ mouse (second panel from top), and a newborn St14–/– mouse (second panel from bottom). Skin extracts from a newborn mouse expressing zymogen-locked (Prss8zym/zym) endogenous prostasin is included as reference (bottom panel). f Ratio of activated prostasin to total prostasin in skin extracts from newborn St14zym/zym (left bar, n = 7), St14+/+ (middle bar, n = 7), and St14–/– (right bar, n = 2) mice, quantified as in (e). Data are shown as mean ± SD. *P = 0.0011 was determined by one-way ANOVA, two-tailed. Additional file 1: Raw supporting data Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/28571576), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, CyTOF-ready, Flow
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

Matriptase/ST14 Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human Matriptase/ST14 Catalytic Domain
  Val615-Val855
  Accession # Q9Y5Y6
• Specificity: Detects human Matriptase/ST14 Catalytic Domain in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: ST14
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Intracellular Staining by Flow Cytometry 2.5 ug/10^6 cells
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Matriptase/ST14 Antibody [Unconjugated]

• EC 3.4.21
• Epithin
• HAI
• Matriptase
• Membrane-type serine protease 1
• MTSP1
• MT-SP1EC 3.4.21.109
• prostamin
• PRSS14
• Serine protease 14
• Serine protease TADG-15
• SNC19
• SNC19MTSP1
• ST14
• suppression of tumorigenicity 14 (colon carcinoma)
• suppression of tumorigenicity 14 (colon carcinoma, matriptase, epithin)
• suppressor of tumorigenicity 14 protein
• TADG15
• TADG-15
• TMPRSS14
• tumor associated differentially expressed gene 15 protein
• Tumor-associated differentially-expressed gene 15 protein

Background

Human matriptase, encoded by the ST14 (suppression of tumorogenicity 14) gene, is also known as tumor associated differentially expressed gene 15 protein/TADG‑15), epithin, and membrane-type serine protease 1/MT-SP1 (1). Predicted to have a significant role in tumor biology, matriptase may be a novel target for anti-cancer therapy (2). However, expressed in most human epithelia, matriptase is also important in several physiological processes (1). For example, it activates prostasin to initiate a protease cascade that is essential for epidermal differentiation (3), and it converts a single-chain IGFBP-rp1 into the two-chain form (4).

Matriptase is a type II transmembrane serine protease with a complex modular structure (1). The 855 amino acid (aa) sequence of human matriptase consists of a cytoplasmic tail (aa 1-55), a transmembrane domain (aa 56-76), and an extracellular portion (aa 77-855). The latter contains the following domains: SEA (aa 86-201), two CUBs (aa 214-334 and 340-447), four LDLRAs (aa 452-486, 487-523, 524-560, and 566-603), and a serine protease (aa 615-855). The physiological activation of the single-chain zymogen requires the cleavage at the SEA domain within the ER or Golgi, association with HAI-1, which facilitates the transport of the protease to the cell surface, and auto-cleavage at QAR-V(615)VGG (1). The activated matriptase is inhibited by HAI-1, and the resulting HAI-1 complex can be shed from the cell surface (1). R&D Systems rhST14 corresponds to the catalytic domain, and is inhibited effectively by rhHAI-1 and rhHAI-2A (R&D Systems, Catalog # 1048-PI and 1106-PI).

1. List, K. et al. (2006) Mol. Med. 12:1.
2. Uhland, K. (2006) Cell. Mol. Life Sci. 63:2968.
3. Netzel-Arnett, S. et al. (2006) J. Biol. Chem. 281:32941.
4. Ahmed, S. et al. (2006) FEBS J. 273:615.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Matriptase/ST14 Antibody (AF3946)(18)

Publications using AF3946

• Ehret, E;Stroh, S;Auberson, M;Ino, F;Jäger, Y;Maillard, M;Szabo, R;Bugge, TH;Frateschi, S;Hummler, E; Kidney-Specific Membrane-Bound Serine Proteases CAP1/Prss8 and CAP3/St14 Affect ENaC Subunit Abundances but Not Its Activity Cells 2023-09-23 [PMID: 37830556] (Western Blot, Mouse)
• Li-Mei Chen, Karl X. Chai Exosome-Mediated Activation of the Prostasin-Matriptase Serine Protease Cascade in B Lymphoma Cells Cancers (Basel) 2023-07-28 [PMID: 37568664]
• Daniel Essigke, Alexandr V. Ilyaskin, Matthias Wörn, Bernhard N. Bohnert, Mengyun Xiao, Christoph Daniel et al. Zymogen‐locked mutant prostasin (Prss8) leads to incomplete proteolytic activation of the epithelial sodium channel (ENaC) and severely compromises triamterene tolerance in mice Acta Physiol (Oxf) 2021-05-01 [PMID: 33650216]
• CJ Wu, M Lu, X Feng, G Nakato, MC Udey Matriptase Cleaves EpCAM and TROP2 in Keratinocytes, Destabilizing Both Proteins and Associated Claudins Cells, 2020-04-21;9(4):. 2020-04-21 [PMID: 32326212] (Western Blot, Human)
• L Drees, T Königsmann, MHJ Jaspers, R Pflanz, D Riedel, R Schuh Conserved function of the matriptase-prostasin proteolytic cascade during epithelial morphogenesis PLoS Genet., 2019-01-02;15(1):e1007882. 2019-01-02 [PMID: 30601807] (IHC, Drosophila)
• S Friis, D Tadeo, SM Le-Gall, HJ Jürgensen, KU Sales, E Camerer, TH Bugge Matriptase zymogen supports epithelial development, homeostasis and regeneration BMC Biol., 2017-06-01;15(1):46. 2017-06-01 [PMID: 28571576] (IHC, Western Blot, Mouse)
• MS Buzza, TA Johnson, GD Conway, EW Martin, S Mukhopadhy, T Shea-Donoh, TM Antalis Inflammatory cytokines down-regulate the barrier protective prostasin-matriptase proteolytic cascade early in experimental colitis J. Biol. Chem., 2017-05-10;0(0):. 2017-05-10 [PMID: 28490634] (IHC, Mouse)
• Chuan-Jin Wu, Xu Feng, Michael Lu, Sohshi Morimura, Mark C. Udey Matriptase-mediated cleavage of EpCAM destabilizes claudins and dysregulates intestinal epithelial homeostasis Journal of Clinical Investigation 2017-02-01 [PMID: 28094766]
• S Friis, DH Madsen, TH Bugge Distinct Developmental Functions of Prostasin (CAP1/PRSS8) Zymogen and Activated Prostasin J. Biol. Chem, 2015-12-30;291(6):2577-82. 2015-12-30 [PMID: 26719335] (Simple Western, Western Blot, Mouse)
• Zoratti G, Tanabe L, Varela F, Murray A, Bergum C, Colombo E, Lang J, Molinolo A, Leduc R, Marsault E, Boerner J, List K Targeting matriptase in breast cancer abrogates tumour progression via impairment of stromal-epithelial growth factor signalling. Nat Commun, 2015-04-15;6(0):6776. 2015-04-15 [PMID: 25873032] (IHC-P, Western Blot, Human, Mouse)
• Sales K, Friis S, Abusleme L, Moutsopoulos N, Bugge T Matriptase promotes inflammatory cell accumulation and progression of established epidermal tumors. Oncogene, 2014-12-08;34(35):4664-72. 2014-12-08 [PMID: 25486433] (IHC-P, Western Blot, Mouse)
• Szabo R, Peters D, Kosa P, Camerer E, Bugge T Regulation of feto-maternal barrier by matriptase- and PAR-2-mediated signaling is required for placental morphogenesis and mouse embryonic survival. PLoS Genet, 2014-07-31;10(7):e1004470. 2014-07-31 [PMID: 25078604] (IHC-P, Western Blot, Mouse)
• Beaulieu A, Gravel E, Cloutier A, Marois I, Colombo E, Desilets A, Verreault C, Leduc R, Marsault E, Richter M Matriptase proteolytically activates influenza virus and promotes multicycle replication in the human airway epithelium. J Virol, 2013-01-30;87(8):4237-51. 2013-01-30 [PMID: 23365447] (ICC, Human)
• Szabo , Roman, Uzzun Sales , Katiuchi, Kosa , Peter, Shylo , Natalia, Godiksen , Sine, Hansen , Karina K, Friis , Stine, Gutkind , J Silvio, Vogel , Lotte K, Hummler , Edith, Camerer , Eric, Bugge , Thomas H Reduced prostasin (CAP1/PRSS8) activity eliminates HAI-1 and HAI-2 deficiency-associated developmental defects by preventing matriptase activation. PLoS Genet, 2012-08-30;8(8):e1002937. 2012-08-30 [PMID: 22952456] (IHC-P, Western Blot, Mouse)
• Szabo R, Rasmussen AL, Moyer AB c-Met-induced epithelial carcinogenesis is initiated by the serine protease matriptase. Oncogene, 2011-01-10;30(17):2003-16. 2011-01-10 [PMID: 21217780] (IHC-P, Western Blot, Mouse)
• Fang JD, Chou HC, Tung HH Endogenous expression of matriptase in neural progenitor cells promotes cell migration and neuron differentiation. J. Biol. Chem., 2010-12-13;286(7):5667-79. 2010-12-13 [PMID: 21149451] (ICC, IHC-P, Western Blot, Mouse)
• Sales KU, Masedunskas A, Bey AL Matriptase initiates activation of epidermal pro-kallikrein and disease onset in a mouse model of Netherton syndrome. Nat. Genet., 2010-07-25;42(8):676-83. 2010-07-25 [PMID: 20657595] (IHC, Mouse)
• Szabo R, Kosa P, List K, Bugge TH Loss of matriptase suppression underlies spint1 mutation-associated ichthyosis and postnatal lethality. Am. J. Pathol., 2009-04-23;174(6):2015-22. 2009-04-23 [PMID: 19389929] (Western Blot, Mouse)

Bioinformatics

• Gene Symbol: ST14
• Uniprot:
  • Q9Y5Y6()