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Purified anti-mouse/human PNAd Antibody

Pricing & Availability
Clone
MECA-79 (See other available formats)
Regulatory Status
RUO
Other Names
PNAd, Peripheral Node Addressin, MECA-79
Isotype
Rat IgM, κ
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Product Citations
publications
A_MECA-79_PURE_PNAd_IHCF_090718
C57BL/6 mouse frozen lymph node section was fixed with 4% paraformaldehyde(PFA) for 10 minutes at room temperature and blocked with 5% FBS for 30 minutes at room temperature. Then the section was stained with 10 µg/ml of PNAd (clone MECA-79) purified, 5 µg/ml of CD45R/B220 (clone RA3-6B2) Alexa Fluor® 647 (green) overnight at 4°C, followed by 5 µg/ml of anti-rat IgM (clone MRM-47) Alexa Fluor® 594 (red) for 2 hours at room temperature. Nuclei were counterstained with DAPI (blue). The image was captured with a 10X objective.
  • A_MECA-79_PURE_PNAd_IHCF_090718
    C57BL/6 mouse frozen lymph node section was fixed with 4% paraformaldehyde(PFA) for 10 minutes at room temperature and blocked with 5% FBS for 30 minutes at room temperature. Then the section was stained with 10 µg/ml of PNAd (clone MECA-79) purified, 5 µg/ml of CD45R/B220 (clone RA3-6B2) Alexa Fluor® 647 (green) overnight at 4°C, followed by 5 µg/ml of anti-rat IgM (clone MRM-47) Alexa Fluor® 594 (red) for 2 hours at room temperature. Nuclei were counterstained with DAPI (blue). The image was captured with a 10X objective.
  • B_MECA-79_PURE_PNAd_IHCP_011316
    Human paraffin-embedded tonsil tissue slices were prepared with a standard protocol of deparaffination and rehydration. Antigen retrieval was done with Tris-Buffered Saline 1X (1.0M, pH7.4) at 95°C for 40 minutes. Tissue was washed with PBS/ 0.05% Tween20 twice for five minutes and blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the tissue was stained with 10 µg/ml of purified anti-mouse/human PNAd antibody (clone MECA-79) at 4°C overnight. On the next day, the tissue was washed twice with PBS and stained with 2 µg/ml of biotin conjugated mouse anti-rat IgM secondary antibody for one hour at room temperature. After the incubation, the tissue was washed twice with PBS for five minutes and stained with Alexa Fluor® 488 Streptavidin (green) for 30 minutes at room temperature. Nuclei were counterstained with DAPI (blue). The image was captured with a 10X objective.
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120801 50 µg £49
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120802 500 µg £180
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Description

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 β1,3-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.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Mouse, Human
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Mouse lymph node stromal cells
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

IHC-F - Quality tested
IHC-P - Verified
FC, IP, WB - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by immunohistochemical staining on frozen tissue sections. For immunohistochemistry, a concentration range of 2.5 - 10 µg/ml is suggested. For immunohistochemical staining on formalin-fixed paraffin-embedded tissue sections, the suggested use of this reagent is 5.0 - 10 µg/ml. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Additional reported applications (for the relevant formats) include: in vitro and in vivo blocking1-4 of cell adhesion, immunohistochemical staining1,2,6 of acetone-fixed frozen sections and 4% paraformaldehyde-fixed paraffin-embedded sections, flow cytometry10,11, immunoprecipitation3,5, and Western blotting6,11.

Application References

(PubMed link indicates BioLegend citation)
  1. Streeter PR, et al. 1988. J. Cell Biol. 107:1853. (IHC, Block)
  2. Michie SA, et al. 1993. Amer. J. Path. 143:1688. (IHC, Block)
  3. Berg EL, et al. 1991. J. Cell Biol. 114:343. (IP, Block)
  4. Berg EL, et al. 1993. Nature 366:695. (Block)
  5. Hemmerich S, et al. 1994. J. Exp. Med. 180:2219. (IP)
  6. Samulowitz U, et al. 2002. Am. J. Path. 160:1669. (IHC, WB)
  7. Burke SD, et al. 2007. Diabetes 56:2919. PubMed
  8. Hirakawa J, et al. 2010. J. Biol. Chem. 285:40864. PubMed
  9. Thomas SN, et al. 2012. J. Immunol. 189:2181. PubMed.
  10. Izawa D, et al. 1999. Int. Immunol. 11(12):1989-98. (FC, ICC)
  11. Sinha RK, et al. 2006. Immunology. 119(4):461-9. (FC, IHC-F, WB)
Product Citations
  1. Sawada J, et al. 2022. Cancer Immunol Res. 10:468. PubMed
  2. Li K, et al. 2022. Cancer Cell. 40:1374. PubMed
  3. Kozlovski S, et al. 2023. Front Immunol. 13:1041552. PubMed
  4. Hirakawa J, et al. 2010. J Biol Chem. 285:40864. PubMed
  5. Nagy D, et al. 2021. Commun Biol. 4:993. PubMed
  6. Menzel L, et al. 2021. Cell Rep. 37:109878. PubMed
  7. Thomas S, et al. 2012. J Immunol. 189:2181. PubMed
  8. Menzel L, et al. 2022. STAR Protoc. 3:101267. PubMed
  9. Li Q, et al. 2020. Int J Oral Sci. 12:24. PubMed
  10. Vito A, et al. 2021. Commun Biol. 4:859. PubMed
  11. Noort A, et al. 2015. Am J Pathol. 185: 1935-1943. PubMed
  12. Burke S, et al. 2007. Diabetes. 56:2919. PubMed
  13. Jiao S, et al. 2022. J Immunother Cancer. 10:. PubMed
  14. Lynskey N, et al. 2015. PLoS One. 11: 1005137. PubMed
RRID
AB_493554 (BioLegend Cat. No. 120801)
AB_493554 (BioLegend Cat. No. 120802)

Antigen Details

Structure
Sulphated CD34, GlyCAM-1 and MAdCAM-1, Sgp200, sialomucin
Distribution

High endothelial venules (HEV) in lymphoid tissues, inflamed tissues and rheumatoid synovia

Function
Leukocyte homing, rolling
Ligand/Receptor
CD62L
Biology Area
Cell Adhesion, Cell Biology, Immunology
Molecular Family
Adhesion Molecules
Antigen References

1. Streeter PR, et al. 1988. J. Cell Biol. 107:1853.
2. Berg EL, et al. 1993. Nature 366:695.
3. Hemmerich S, et al. 1994. J. Exp. Med. 180:2219.
4. Mitoma J, et al. 2003. J. Biol. Chem. 278:9953.
5. Bruehl RE, et al. 2000. J. Biol. Chem. 275:32642.
6. Bistrup A, et al. 2004. Am. J. Pathol. 164:1635

Gene ID
123803 View all products for this Gene ID 18203 View all products for this Gene ID
UniProt
View information about PNAd on UniProt.org
Go To Top Version: 4    Revision Date: 09/07/2018

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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