Peripheral Node Addressin Antibody (MECA-79) - Non-Recombinant Monoclonal

Immunohistochemistry: Peripheral Node Addressin Antibody (MECA-79) - Non-Recombinant Monoclonal [NB100-77673] - Local release of anti-IL-6 from IMB suppresses LN fibrosis. The skin allografts were harvested at 7 days post-transplantation. (A-i) Lymphatic vessel expansion (Lyve-1+) and HEV elongation (MECA79+) were similar between all groups. (A-ii,iii) Dense staining of collagen I 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.

Western Blot: Peripheral Node Addressin Antibody (MECA-79) - Non-Recombinant Monoclonal [NB100-77673] - 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. Image using the standard format of this product.

Western Blot: Peripheral Node Addressin Antibody (MECA-79) - Non-Recombinant Monoclonal [NB100-77673] - Total protein from Human tonsil (lane 1) and human lymph node (lane 2) 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 ug/mL anti-PNAd in 1% milk, and detected with an anti-rat IgM HRP conjugated secondary antibody using chemiluminescence.

Immunohistochemistry-Paraffin: Peripheral Node Addressin Antibody (MECA-79) - Non-Recombinant Monoclonal [NB100-77673] - 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: Peripheral Node Addressin Antibody (MECA-79) - Non-Recombinant Monoclonal [NB100-77673] - 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. Image using the standard format of this product.

Immunocytochemistry/ Immunofluorescence: Peripheral Node Addressin Antibody (MECA-79) - Non-Recombinant Monoclonal [NB100-77673] - Local release of anti-IL-6 from IMB suppresses LN fibrosis. The skin allografts were harvested at 7 days post-transplantation. (A-i) Lymphatic vessel expansion (Lyve-1+) & HEV elongation (MECA79+) were similar between all groups. (A-ii,iii) Dense staining of collagen I & PDPN was seen in DLNs harvested from control & GelMA group compared to those from GelMA/anti-IL-6 group. (representative images from 4 different mice per group). Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31024011), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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

Peripheral Node Addressin Antibody (MECA-79) - Non-Recombinant Monoclonal Summary

• Immunogen: Mouse lymph node stromal cells
• Isotype: IgM Kappa
• Clonality: Monoclonal
• Host: Rat
• Purity: IgM purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence reported in scientific literature (PMID 31024011)
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Frozen 1:10-1:500
  • Immunohistochemistry-Paraffin 1:10-1:500
  • Immunoprecipitation 1:10-1:500
  • In vivo assay reported in scientific literature (PMID 30277476)
  • 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 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: IgM purified

Alternate Names for Peripheral Node Addressin Antibody (MECA-79) - Non-Recombinant Monoclonal

• 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 (NB100-77673)(23)

Publications using NB100-77673

• Gavish A, Tyler M, Greenwald AC et al. Hallmarks of transcriptional intratumour heterogeneity across a thousand tumours Nature 2023-06-01 [PMID: 37258682]
• Wang Z, Shi X, Zhao Y et al. DC101, an anti-VEGFR2 agent, promotes high-endothelial venule formation and immune infiltration versus SAR131675 and fruquintinib Biochemical and biophysical research communications 2023-06-18 [PMID: 37084488] (IHC, ICC/IF)
• 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 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)
• Goveia J, Rohlenova K, Taverna F et al. An Integrated Gene Expression Landscape Profiling Approach to Identify Lung Tumor Endothelial Cell Heterogeneity and Angiogenic Candidates Cancer Cell 2020-01-13 [PMID: 31935371]
• 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.
• 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.

FAQs for Peripheral Node Addressin Antibody (NB100-77673). (Showing 1 - 3 of 3 FAQs).

1. What is the recommended concentration for the use of this antibody on human paraffin sections, as well as a specific protocol for this antibody?
   • We do not have a specific protocol for this product, and we recommend a starting dilution of 1:100-1:200 for IHC-P. We performed heat induced antigen retrieval with Citrate buffer pH6.0.
2. We are very interested in your MECA-79 antibody. The catalog# is NB100-77673. We want to inject the antibody to mouse as blocking antibody. Our question is whether the antibody solution contains toxic sodium azide? Thanks.
   • NB100-77673 does contain 0.09% sodium azide. We do offer antibody cleanup kits which you might find useful as they remove unwanted substances such as sodium azide.
3. I am searching for an antibody against MECA 79 that reacts with human, mouse primary cells for WB or IP-WB. I did use one other vendor's, but it hard for me to use that for WB. Since my project will use MECA 79 antibody a lot, so I found today that you have this antibody. I would like to try it. My question is: Do you have the image picture for WB?
   • Unfortunately, we do not have a western blot image of this product. We do have a publication that used this antibody in western blot among other immunoassays. It is currently not available on our website, but you may view it using this link: PMC3003387. We also do not have a smaller size of this antibody available. I am sorry for any inconvenience. We will fully guarantee all listed applications and species.