Trypsin Pan Specific/PRSS1/2/3 Antibody [Unconjugated]

Western blot shows lysates of human pancreas. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Trypsin Pan Specific (PRSS1/2/3) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3586) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for Trypsin at approximately 24 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Western blot shows recombinant human Trypsin 1, 2, and 3. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Trypsin Pan Specific (PRSS1/2/3) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3586) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for Trypsin at approximately 24 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Trypsin was detected in immersion fixed paraffin-embedded sections of human pancreas using Sheep Anti-Human Trypsin Pan Specific (PRSS1/2/3) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3586) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to exocrine cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Trypsin was detected in immersion fixed paraffin-embedded sections of human pancreatic cancer tissue using Sheep Anti-Human Trypsin Pan Specific (PRSS1/2/3) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3586) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm of cancer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Human Trypsin 2/PRSS2 was immunoprecipitated from 500 μg of human pancreas lysates with 12.5 ug Mouse Anti-Human Trypsin 2/PRSS2 Monoclonal Antibody (Catalog # MAB3586). The Trypsin 2/PRSS2-antibody complexes were absorbed using Protein G Sepharose. Immunoprecipitated human Trypsin 2/PRSS2 was detected by Western blot using 2 µg/mL of Sheep Anti-Human Trypsin Pan Specific (PRSS1/2/3) Antigen Affinity-purified Polyclonal Antibody (AF3586) under non-reducing conditions and using Western Blot Buffer Group 1.

• Reactivity: Hu
• Applications: WB, IHC, IP, B/N
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

Trypsin Pan Specific/PRSS1/2/3 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Trypsin 2
  Ala16-Ser247
  Accession # P07478
• Specificity: Detects human Trypsin Pan Specific (PRSS1/2/3) in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Neutralization 0.3 ug/mL
  • Western Blot 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. *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.

Notes

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

Alternate Names for Trypsin Pan Specific/PRSS1/2/3 Antibody [Unconjugated]

• TRY2
• TRY8
• TRYP2
• Trypsin

Background

Trypsin is a general term for any of three 24 kDa gene products that belong to the peptidase S1 family of enzymes. Trypsin in Greek means “rubbing or friction”, and it was chosen here because the first trypsins were extracted from pancreas via a glycerin-based rubbing maceration. Trypsin-1 (cationic), -2 (anionic) , and -3 (mesotrypsin) are synthesized as 26 kDa trypsinogens (plus a 35 kDa trypsinogen-3 isoform) that are 247 amino acids (aa) in length. The first 15 aa constitute a signal sequence, followed by an enterokinase-cleavable eight aa propeptide, and a 224 aa mature molecule. Asp194 is linked to enzyme activity, and Tyr154 is sulfated. Over their mature regions, the three trypsins share 84% aa identity.  Mouse trypsin-1 shares 74% aa identity with the human trypsin consensus sequence. Trypsin-1 and -2 cleave peptide bonds carboxylterminal to a Lys or Arg. Trypsin-3 has limited activity on trypsin inhibitors, and thus regulates general trypsin activity. Trypsin-1, -2, and -3 are expressed by multiple tissues, including pancreas (for general digestion), liver, lung and skin. In blood, trypsinogen-1 and -2 circulate at ng/mL concentrations. In pancreatic fluid, trypsin-1, -2, and -3 are found in a 58:40:2 ratio. This antibody did not neutralize the activity of rhTrypsin 1 and rhTrypsin 3 significantly.  

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 Trypsin Pan Specific/PRSS1/2/3 Antibody (AF3586)(4)

Publications using AF3586

• Y Aghazadeh, F Poon, F Sarangi, FTM Wong, ST Khan, X Sun, R Hatkar, BJ Cox, SS Nunes, MC Nostro Microvessels support engraftment and functionality of human islets and hESC-derived pancreatic progenitors in diabetes models Cell Stem Cell, 2021-09-03;0(0):. 2021-09-03 [PMID: 34480863] (IHC, Human)
• P Augsornwor, KG Maxwell, L Velazco-Cr, JR Millman Single-Cell Transcriptome Profiling Reveals &beta Cell Maturation in Stem Cell-Derived Islets after Transplantation Cell Rep, 2020-08-25;32(8):108067. 2020-08-25 [PMID: 32846125] (ICC, Human)
• NJ Hogrebe, P Augsornwor, KG Maxwell, L Velazco-Cr, JR Millman Targeting the cytoskeleton to direct pancreatic differentiation of human pluripotent stem cells Nat. Biotechnol., 2020-02-24;0(0):. 2/24/2020 [PMID: 32094658]
• Nostro M, Sarangi F, Yang C, Holland A, Elefanty A, Stanley E, Greiner D, Keller G Efficient generation of NKX6-1+ pancreatic progenitors from multiple human pluripotent stem cell lines. Stem Cell Reports, 2015-04-02;4(4):591-604. 2015-04-02 [PMID: 25843049] (IHC-P, Human)

Reviews for Trypsin Pan Specific/PRSS1/2/3 Antibody (AF3586) (2)

Average Rating: 5

(Based on 2 reviews)

We have 2 reviews tested in 2 species: Mouse, Human and Rat.

reviewed by: Scott Messenger ICC Mouse 06/09/2017

Summary

• Application: Immunocytochemistry
• Sample Tested: Mouse pancreas in OCT
• Species: Mouse

Comments

• Comments: cryosection of mouse pancreas, primary 1/100, secondary 1/200 488. Epi-fluorescence 60 x. signal weak

reviewed by: Scott Messenger WB Human and Rat 05/24/2017

Summary

• Application: Western Blot
• Sample Tested: AR42J rat acinar-like cells,human acini,rat acini
• Species: Human and Rat

Comments

• Comments: 1/1000, 12% gel, 50 ug protein

Bioinformatics

• Uniprot:
  • P07478()