Aminopeptidase PILS/ARTS1 Antibody [Unconjugated]

Western blot shows lysates of COLO 205 human colorectal adenocarcinoma cell line and HDLM-2 human Hodgkin's lymphoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Aminopeptidase PILS/ARTS1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2334) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Aminopeptidase PILS/ARTS1 at approximately 100 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Aminopeptidase PILS/ARTS1 was detected in immersion fixed A549 human lung carcinoma cell line using Goat Anti-Human Aminopeptidase PILS/ARTS1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2334) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Aminopeptidase PILS/ARTS1 was detected in immersion fixed paraffin-embedded sections of human liver using Goat Anti-Human Aminopeptidase PILS/ARTS1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2334) at 1 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). Specific staining was localized to hepatocyte cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Simple Western lane view shows lysates of COLO 205 human colorectal adenocarcinoma cell line and HDLM‑2 human Hodgkin’s lymphoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Aminopeptidase PILS/ARTS1 at approximately 115 kDa (as indicated) using 10 µg/mL of Goat Anti-Human Aminopeptidase PILS/ARTS1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2334). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

• Reactivity: Hu
• Applications: WB, Simple Western, IHC, IP, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Aminopeptidase PILS/ARTS1 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Aminopeptidase PILS/ARTS1
  Ala37-Met941
  Accession # EAW96077
• Specificity: Detects human Aminopeptidase PILS/ARTS1 in direct ELISAs and Western blots. In direct ELISAs, approximately 45% cross‑reactivity with recombinant mouse ARTS1 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: ERAP1
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 1-15 ug/mL
  • Immunohistochemistry 1-15 ug/mL
  • Immunoprecipitation 25 ug/mL
  • Simple Western 10 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 Aminopeptidase PILS/ARTS1 Antibody [Unconjugated]

• Adipocyte-derived leucine aminopeptidase
• A-LAP
• A-LAPALAP
• Aminopeptidase PILS
• ARTS1
• ARTS1aminopeptidase regulator of TNFR1 shedding
• ARTS-1PILSAP
• EC 3.4.11
• EC 3.4.11.-
• EC 3.4.11.1
• endoplasmic reticulum aminopeptidase 1
• endoplasmic reticulum aminopeptidase associated with antigen processing
• ERAAP1
• ERAP1
• KIAA0525APPILS
• PILS-AP
• PILS-APERAAP
• Puromycin-insensitive leucyl-specific aminopeptidase
• Type 1 tumor necrosis factor receptor shedding aminopeptidase regulator

Background

The name of Aminopetidase PILS (Puromycin-Insensitive Leucyl-Specific) describes the two basic properties of this zinc metalloprotease in vitro (1). Also known as ALAP (Adipocyte-derived Leucin AiminoPeptidase), type 1 tumor necrosis factor receptor (TNFR) shedding aminopeptidase regulator and ER aminopeptidase ERAP1 or ERAAP, it is encoded by the ARTS1 gene (2‑4). Aminopeptidase PILS has been identified to regulate antigen presentation, promote TNFR1 ectodomain shedding and associate with hypertension (2‑5).

1. Schomburg, L. (2004) in Handbook of Proteolytic Enzymes (ed. Barrett, et al.) pp. 311, Academic Press, San Diego.
2. Cui, X. et al. (2002) J. Clin. Invest. 110:515.
3. York, I.A. et al. (2002) Nat. Immunol. 3:1177.
4. Serwold, T. et al. (2002) Nature 419:480.
5. Yamamoto, N. et al. (2002) Hum. Mutat. 19:251.

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 Aminopeptidase PILS/ARTS1 Antibody (AF2334)(11)

Publications using AF2334

• Temponeras, I;Samiotaki, M;Koumantou, D;Nikopaschou, M;Kuiper, JJW;Panayotou, G;Stratikos, E; Distinct modulation of cellular immunopeptidome by the allosteric regulatory site of ER aminopeptidase 1 European journal of immunology 2023-05-03 [PMID: 37134263] (Western Blot, Human)
• I Temponeras, G Stamatakis, M Samiotaki, D Georgiadis, H Pratsinis, G Panayotou, E Stratikos ERAP2 Inhibition Induces Cell-Surface Presentation by MOLT-4 Leukemia Cancer Cells of Many Novel and Potentially Antigenic Peptides International Journal of Molecular Sciences, 2022-02-08;23(3):. 2022-02-08 [PMID: 35163832] (Western Blot, Human)
• Akiko Arakawa, Emma Reeves, Sigrid Vollmer, Yukiyasu Arakawa, Mengwen He, Adrian Galinski et al. ERAP1 Controls the Autoimmune Response against Melanocytes in Psoriasis by Generating the Melanocyte Autoantigen and Regulating its Amount for HLA-C*06:02 Presentation1 The Journal of Immunology 2021-11-01 [PMID: 34580106]
• Wouter J. Venema, Sanne Hiddingh, Joke H. de Boer, Frans H. J. Claas, Arend Mulder, Anneke I. den Hollander et al. ERAP2 Increases the Abundance of a Peptide Submotif Highly Selective for the Birdshot Uveitis-Associated HLA-A29 Frontiers in Immunology 2021-02-25 [PMID: 33717175]
• P Jiang, RN Veenstra, A Seitz, IM Nolte, BG Hepkema, L Visser, A van den Be, A Diepstra Interaction between ERAP Alleles and HLA Class I Types Support a Role of Antigen Presentation in Hodgkin Lymphoma Development Cancers, 2021-01-22;13(3):. 2021-01-22 [PMID: 33499248] (IHC, Western Blot, Human)
• Irma Saulle, Claudia Vanetti, Sara Goglia, Chiara Vicentini, Enrico Tombetti, Micaela Garziano et al. A New ERAP2/Iso3 Isoform Expression Is Triggered by Different Microbial Stimuli in Human Cells. Could It Play a Role in the Modulation of SARS-CoV-2 Infection? Cells 2020-08-24 [PMID: 32847031]
• Despoina Koumantou, Eilon Barnea, Adrian Martin-Esteban, Zachary Maben, Athanasios Papakyriakou, Anastasia Mpakali et al. Editing the immunopeptidome of melanoma cells using a potent inhibitor of endoplasmic reticulum aminopeptidase 1 (ERAP1) Cancer Immunology, Immunotherapy 2019-08-01 [PMID: 31222486]
• Jonas J W Kuiper, Jessicavan van Setten, Matthew Devall, Mircea Cretu-Stancu, Sanne Hiddingh, Roel A Ophoff et al. Functionally distinct ERAP1 and ERAP2 are a hallmark of HLA-A29-(Birdshot) Uveitis Human Molecular Genetics 2018-09-28 [PMID: 30215709]
• A Papakyriak, E Reeves, M Beton, H Mikolajek, L Douglas, G Cooper, T Elliott, JM Werner, E James The partial dissociation of MHC class I bound peptides exposes their N terminus to trimming by endoplasmic reticulum aminopeptidase 1 J. Biol. Chem., 2018-03-29;0(0):. 2018-03-29 [PMID: 29599287] (Western Blot, Human)
• SR Harrison, T Scambler, L Oubussad, C Wong, M Wittmann, MF McDermott, S Savic Inositol-Requiring Enzyme 1-Mediated Downregulation of MicroRNA (miR)-146a and miR-155 in Primary Dermal Fibroblasts across ThreeTNFRSF1AMutations Results in Hyperresponsiveness to Lipopolysaccharide Front Immunol, 2018-02-06;9(0):173. 2018-02-06 [PMID: 29467762] (ICC, Human)
• Emma Reeves, Christopher J. Edwards, Tim Elliott, Edward James Naturally Occurring ERAP1 Haplotypes Encode Functionally Distinct Alleles with Fine Substrate Specificity The Journal of Immunology 2013-07-01 [PMID: 23733883]

Bioinformatics

• Gene Symbol: ERAP1
• Uniprot:
  • EAW96077()