Peripheral Node Addressin Antibody (MECA-79R) - BSA Free

Peripheral Node Addressin was detected in immersion fixed paraffin-embedded sections of human Tonsil using Rat Anti-Human Peripheral Node Addressin (MECA-79R), Monoclonal Antibody (Catalog #NBP2-78792) at 1:50 dilution at 37° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 647 Goat anti-Rat IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647RT) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to endothelial cells. Protocol available in COMET™ Panel Builder.

Western Blot: Peripheral Node Addressin Antibody (MECA-79R) [NBP2-78792] - Total protein from human Tonsil, Lymph node, Spleen and mouse Spleen was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-PNAd in blocking buffer and detected with an anti-rat HRP secondary antibody using West Pico PLUS chemiluminescence detection reagent.

Immunohistochemistry-Paraffin: Peripheral Node Addressin Antibody (MECA-79R) [NBP2-78792] - Analysis of FFPE mouse adipose tissue section (with lymph node areas) using Peripheral Node Addressin antibody (clone MECA-79) at 1:100. The staining was developed with HRP-DAB detection method and the counterstaining was performed using hematoxylin. This Peripheral Node Addressin antibody generated a strong and specific staining of MECA-79 antigen in the the cytoplasm and the membranes of high endothelial venules (HEVs) aka peripheral lymph node addressin (PNAd) in lymph node areas of tested adipose tissue section.

Immunohistochemistry-Paraffin: Peripheral Node Addressin Antibody (MECA-79R) [NBP2-78792] - Analysis of FFPE human tonsil tissue section using Peripheral Node Addressin antibody (clone MECA-79R) at 1:100. The staining was developed with HRP-DAB detection method and the counterstaining was performed using hematoxylin.

Immunohistochemistry: Peripheral Node Addressin Antibody (MECA-79R) [NBP2-78792] - Local release of anti-IL-6 from IMB suppresses LN fibrosis. The skin allografts were harvested at 7 days post-transplantation. Lymphatic vessel expansion (Lyve-1+) and HEV elongation (MECA79+) were similar between all groups. Dense staining of collagen I and PDPN was seen in DLNs harvested from control and GelMA group compared to those from GelMA/anti-IL-6 group. (representative images from 4 different mice per group). Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-019-42349-w), licensed under a CC-BY license.

• Reactivity: Hu, Mu
• Applications: WB, Func, ICC/IF, IHC, mIF
• Clone: MECA-79R
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

Peripheral Node Addressin Antibody (MECA-79R) - BSA Free Summary

• Description: Clone MECA-79R (rat IgG1) is a recombinant version of the original clone MECA-79 (rat IgM).
• Immunogen: Mouse lymph node stromal cells
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Rat
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence reported in scientific literature (PMID 31278331)
  • Immunohistochemistry 1:100-1:500
  • Immunohistochemistry-Frozen 1:100-1:500
  • Immunohistochemistry-Paraffin 1:100-1:500
  • In vivo assay reported in scientific literature (PMID 30277476)
  • Multiplex Immunofluorescence 1:50
  • Western Blot 1:100-1:2000
• Application Notes: Additional reported application of in vitro and in vivo blocking of cell adhesion.

Packaging, Storage & Formulations

• Storage: Store at -20 °C.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein A or G purified

Alternate Names for Peripheral Node Addressin Antibody (MECA-79R) - BSA Free

• MECA-79
• Peripheral Node Addressin
• PNAd

Background

MECA-79 recognizes a carbohydrate epitope shared with a group of sulfation decorated sialomucins, including sulfated ligands for CD62L (CD34, GlyCAM-1, Sgp200, and a subset of MAdCAM-1). This set antigens has been referred to as peripheral node addressin (PNAd) with the molecular mass 50-250 kD. It has been identified that GlcNAc-6-SO4 sulfation contributes to MECA-79 binding and the core 1 beta1,3-N-acetylglucosaminyltransferase is required for the generation of the MECA-79 epitope. MECA-79 is expressed on high endothelial venules (HEV) of lymphoid tissues, chronic inflamed tissues and rheumatoid synovia. The interaction of PNAd with CD62L receptor is involved in tethering and rolling of lymphocytes along HEV in lymphoid tissues. MECA-79 antibody reacts with mouse, human and many other species PNAd and blocks L-selectin-dependent lymphocyte adhesion in vitro and in vivo.

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 Peripheral Node Addressin Antibody (NBP2-78792)(26)

Publications using NBP2-78792

• Gavish A, Tyler M, Greenwald AC et al. Hallmarks of transcriptional intratumour heterogeneity across a thousand tumours Nature 2023-06-01 [PMID: 37258682]
• Park JA, Espinosa-Cotton M, Guo HF et al. Targeting tumor vasculature to improve antitumor activity of T cells armed ex vivo with T cell engaging bispecific antibody Journal for immunotherapy of cancer 2023-03-01 [PMID: 36990507] (IHC, Mouse)
• Quigley LT, Pang L, Tavancheh E et al. Protocol for investigating tertiary lymphoid structures in human and murine fixed tissue sections using Opal�-TSA multiplex immunohistochemistry STAR protocols 2023-01-10 [PMID: 36633948] (IHC-P, Human)
  
  Details:
  Dilution used in IHC-P 1:100
• Zhao J, Jung S, Li X et al. Delivery of costimulatory blockade to lymph nodes promotes transplant acceptance in mice Journal of Clinical Investigation 2022-12-15 [PMID: 36519543]
• Smith KJ, Minns D, McHugh BJ Et al. The antimicrobial peptide cathelicidin drives development of experimental autoimmune encephalomyelitis in mice by affecting Th17 differentiation PLoS Biol 2022-08-26 [PMID: 36026478]
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Li Y, Amaladas N, O'Mahony M et al. Treatment with a VEGFR-2 antibody results in intra-tumor immune modulation and enhances anti-tumor efficacy of PD-L1 blockade in syngeneic murine tumor models PloS one 2022-07-18 [PMID: 35849586] (IHC-P, Mouse)
• Pinkard, H, Baghdassarian, H Et al. Learned adaptive multiphoton illumination microscopy for large-scale immune response imaging. Nat Commun 2021-03-26 [PMID: 33772022]
• Li X, Zhao J, Kasinath V et al. Lymph node fibroblastic reticular cells deposit fibrosis-associated collagen following organ transplantation J. Clin. Invest. 2020-06-29 [PMID: 32597832] (IHC-Fr, Human)
  
  Details:
  Citation using the Non-Recombinant Monoclonal format of this antibody.
• He B, Johansson-Percival A, Backhouse J et al. Remodeling of Metastatic Vasculature Reduces Lung Colonization and Sensitizes Overt Metastases to Immunotherapy Cell Rep. [PMID: 31968248] (ICC/IF, Mouse, Human)
• Helmink BA, Reddy SM, Gao J et al. B cells and tertiary lymphoid structures promote immunotherapy response Nature. [PMID: 31942075] (MI, Human)
• Matsubara S, Seki M, Suzuki S et al. Tertiary lymphoid organs in the inflammatory myopathy associated with PD-1 inhibitors J Immunother Cancer. 2019-09-18 [PMID: 31533865] (IF/IHC, Human)
  
  Details:
  Citation used the Non-Recombinant Monoclonal format of this antibody.
• Li X, Bean A, Uehara M et al. Immune heterogeneity of head and tail pancreatic lymph nodes in non-obese diabetic mice Sci Rep 2019-07-05 [PMID: 31278331] (ICC/IF)
• Uehara M, Li X, Sheikhi A et al. Anti-IL-6 eluting immunomodulatory biomaterials prolong skin allograft survival. Sci Rep. 2019-04-25 [PMID: 31024011] (ICC/IF, Mouse)
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Bahmani B, Uehara M, Ordikhani F et al Ectopic high endothelial venules in pancreatic ductal adenocarcinoma: A unique site for targeted delivery. EBioMedicine. 2018-12-01 [PMID: 30497977] (IHC-Fr, Human)
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Bahmani B, Uehara M, Jiang L et al Targeted delivery of immune therapeutics to lymph nodes prolongs cardiac allograft survival. J Clin Invest. 2018-11-01 [PMID: 30277476] (In Vivo, Mouse)
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Hautz T, Zelger BG, Nasr IW et al Lymphoid neogenesis in skin of human hand, nonhuman primate, and rat vascularized composite allografts. Transpl Int. 2014-09-01 [PMID: 24853399] (IHC-P, Human)
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Hirakawa J, Tsuboi K, Sato K et al Novel anti-carbohydrate antibodies reveal the cooperative function of sulfated N- and O-glycans in lymphocyte homing. J Biol Chem. 2010-12-24 [PMID: 20929857]
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Burke SD, Dong H, Hazan AD et al Aberrant endometrial features of pregnancy in diabetic NOD mice Diabetes. 2007-09-01 [PMID: 17827401]
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Drayton DL, Ying X, Lee J et al Ectopic LT alpha beta directs lymphoid organ neogenesis with concomitant expression of peripheral node addressin and a HEV-restricted sulfotransferase. J Exp Med. 2003-05-05 [PMID: 12732657] (IF/IHC, Mouse)
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Samulowitz U, Kuhn A, Brachtendorf G et al Human endomucin: distribution pattern, expression on high endothelial venules, and decoration with the MECA-79 epitope. Am J Pathol. 2002-05-01 [PMID: 12000719]
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Csencsits KL, Jutila MA, Pascual DW Nasal-associated lymphoid tissue: phenotypic and functional evidence for the primary role of peripheral node addressin in naive lymphocyte adhesion to high endothelial venules in a mucosal site. J Immunol. 1999-08-01 [PMID: 10415038] (IF/IHC, Mouse)
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Michie SA, Streeter PR, Bolt PA et al The human peripheral lymph node vascular addressin. An inducible endothelial antigen involved in lymphocyte homing. Am J Pathol. 1993-12-16 [PMID: 8256856]
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Hemmerich S, Butcher EC, Rosen SD et al Sulfation-dependent recognition of high endothelial venules (HEV)-ligands by L-selectin and MECA 79, and adhesion-blocking monoclonal antibody. J Exp Med. 1994-12-01 [PMID: 7525849]
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Berg EL, McEvoy LM, Berlin C et al L-selectin-mediated lymphocyte rolling on MAdCAM-1. Nature. 1993-12-16 [PMID: 7505053]
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Streeter PR, Rouse BT, Butcher EC et al Immunohistologic and functional characterization of a vascular addressin involved in lymphocyte homing into peripheral lymph nodes. J Cell Biol. 1988-11-01 [PMID: 2460470]
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.
• Berg EL, Robinson MK, Warnock RA et al The human peripheral lymph node vascular addressin is a ligand for LECAM-1, the peripheral lymph node homing receptor. J Cell Biol. 1991-07-01 [PMID: 1712790]
  
  Details:
  Citation using the Non-Recombinant Monoclonal version of this antibody.