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

Novus Biologicals | Catalog # NB100-77673

Recombinant Monoclonal Antibody
Novus Biologicals
100% Guarantee

Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, In vivo assay

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Immunocytochemistry/ Immunofluorescence, In vivo assay, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Rat IgM Kappa Clone # MECA-79

Format

Non-Recombinant Monoclonal
View all Peripheral Node Addressin Primary Antibodies »

Product Specifications

Immunogen

Mouse lymph node stromal cells

Clonality

Monoclonal

Host

Rat

Isotype

IgM Kappa

Scientific Data Images

Immunohistochemistry: Peripheral Node Addressin Antibody (MECA-79) - Non-Recombinant Monoclonal [NB100-77673]

Immunohistochemistry: Peripheral Node Addressin Antibody (MECA-79) - Non-Recombinant Monoclonal [NB100-77673]

Peripheral-Node-Addressin-Antibody-MECA-79-Non-Recombinant-Monoclonal-Immunohistochemistry-NB100-77673-img0006.jpg
Western Blot: Peripheral Node Addressin Antibody (MECA-79)Non-Recombinant Monoclonal [NB100-77673]

Western Blot: Peripheral Node Addressin Antibody (MECA-79)Non-Recombinant Monoclonal [NB100-77673]

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]

Western Blot: Peripheral Node Addressin Antibody (MECA-79)Non-Recombinant Monoclonal [NB100-77673]

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]

Immunohistochemistry-Paraffin: Peripheral Node Addressin Antibody (MECA-79) - Non-Recombinant Monoclonal [NB100-77673]

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]

Immunohistochemistry: Peripheral Node Addressin Antibody (MECA-79) - Non-Recombinant Monoclonal [NB100-77673]

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

Immunocytochemistry/ Immunofluorescence: Peripheral Node Addressin Antibody (MECA-79) - Non-Recombinant Monoclonal [NB100-77673] -

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 (https://pubmed.ncbi.nlm.nih.gov/31024011), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications

Application
Recommended Usage

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.

Formulation, Preparation, and Storage

Purification

IgM purified

Formulation

PBS

Format

Non-Recombinant Monoclonal

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Peripheral Node Addressin

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.

Alternate Names

MECA-79, Peripheral Node Addressin, PNAd

Additional Peripheral Node Addressin Products

Product Documents

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices

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.

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

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Protocols

View specific protocols for Peripheral Node Addressin Antibody (MECA-79) - Non-Recombinant Monoclonal (NB100-77673):

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer all the time).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs

Showing  1 - 3 of 3 FAQs Showing All

  • Q: 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?

    A: 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.

  • Q: 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.

    A:

    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.

  • Q: What is the recommended concentration for the use of this antibody on human paraffin sections, as well as a specific protocol for this antibody?

    A: 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.

  • Q: 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?

    A: 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.

  • Q: 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.

    A:

    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.

  • Q: What is the recommended concentration for the use of this antibody on human paraffin sections, as well as a specific protocol for this antibody?

    A: 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.

  • Q: 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?

    A: 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.

  • Q: 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.

    A:

    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.

  • Q: What is the recommended concentration for the use of this antibody on human paraffin sections, as well as a specific protocol for this antibody?

    A: 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.

Showing  1 - 3 of 3 FAQs Showing All
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