Overview

  • Product name
    Anti-Aggrecan antibody [6-B-4]
    See all Aggrecan primary antibodies
  • Description
    Mouse monoclonal [6-B-4] to Aggrecan
  • Host species
    Mouse
  • Specificity
    Recognises the linear amino acid sequence 394EPEEPFTFAPEI406 present in the interglobular domain (IGD) of human and bovine aggrecan (in both intact and matrix protease degraded aggrecan) but not pig and several other animals.
  • Tested applications
    Suitable for: ELISA, IHC-P, IHC-Fr, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Cow, Human
  • Immunogen

    Synthetic peptide:

    EPEEPFTFAPEI

    , corresponding to amino acids 413-424 of Human Aggrecan.

  • General notes
    Samples are usually deglycosylated using 0.01 Units Chondroitinase ABC (Sigma), 0.01 Units Keratanase (Seikagaku) and 0.0001 Units Keratanase II (Seikagaku) per 10µg S-GAG of non-deglycosylated aggrecan for optimal epitope recognition in SDS-PAGE and immunohistochemistry – see Little et al. and Caterson et al. Diluted Technomouse culture supernatant.


    This antibody detects aggrecan metabolites (intact or matrix protease-catabolised) in human synovial fluid samples.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: 0.2% Tris HCl
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Primary antibody notes
    This antibody detects aggrecan metabolites (intact or matrix protease-catabolised) in human synovial fluid samples.
  • Clonality
    Monoclonal
  • Clone number
    6-B-4
  • Myeloma
    x63-Ag8.653
  • Isotype
    IgG1
  • Light chain type
    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab3778 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ELISA Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
WB 1/100. The antibody will see a large range of aggrecan metabolites/catabolites ranging from greater than 250kD down to 50-60kDa.
ICC/IF Use at an assay dependent concentration. PubMed: 21192818

Target

  • Relevance
    Aggrecan is a member of a family of large, aggregating proteoglycans (also including versican, brevican and neurocan) which is found in articular cartilage. Aggrecan is composed of three major domains: G1, G2, and G3. Between the G1 and G2 domains there is an interglobulin region (IGD). The IGD region is the major site of cleavage by specific proteases like metalloproteinases (MMPs) and aggrecanase. Aggrecan cleavage has been associated with a number of degenerative diseases including rheumatoid arthritis and osteoarthritis. There is evidence that this family of proteoglycans modulates cell adhesion, migration, and axonal outgrowth in the CNS.
  • Cellular localization
    Secreted; extracellular matrix.
  • Database links
  • Alternative names
    • ACAN antibody
    • AGC 1 antibody
    • AGC1 antibody
    • AGCAN antibody
    • Aggrecan 1 (chondroitin sulfate proteoglycan 1, large aggregating proteoglycan, antigen identified by monoclonal antibody A0122) antibody
    • Aggrecan 1 antibody
    • Aggrecan core protein antibody
    • Aggrecan proteoglycan antibody
    • Aggrecan structural proteoglycan of cartilage antibody
    • Aggrecan1 antibody
    • ATEGQV antibody
    • Cartilage specific proteoglycan core protein antibody
    • Chondroitin sulfate proteoglycan 1 antibody
    • Chondroitin sulfate proteoglycan 1 large aggregating proteoglycan antigen identified by monoclonal antibody A0122 antibody
    • Chondroitin sulfate proteoglycan core protein 1 antibody
    • CSPG 1 antibody
    • CSPG1 antibody
    • CSPGCP antibody
    • JSCATE antibody
    • Large aggregating proteoglycan antibody
    • mcspg antibody
    • mgsk16 antibody
    • MSK 16 antibody
    • MSK16 antibody
    • SEDK antibody
    see all

Images

  • ab3778 at 1/100 staining human articular cartilage by IHC-P. The tissue was formaldehyde fixed and blocked with a serum free protein block. An enzymatic antigen retrieval step was performed and the tissue was then incubated with the antibody for 1 hour. An HRP conjugated goat polyclonal antibody was used as the secondary.

    See Abreview

  • Immunofluorescence analysis of Human articular chondrocytes, staining Aggrecan with ab3778.

    Cells were fixed with paraformaldehyde and blocked with 1% BSA for 1 hour. Cells were incubated with primary antibody overnight at 4°C. An AlexaFluor®488-conjugated rabbit anti-mouse IgG was used as the secondary antibody.
  • ab3778 used at a 1/50 dilution staining Aggrecan in human umbilical cord matrix mesenchymal stromal cells by Immunocytochemistry/ Immunofluorescence.
    Frozen sections of the 3D constructs (10µm) (n=2) were rehydrated with PBS for 5 minutes, and endogenous peroxidase activity was inhibited using 1% hydrogen peroxide in methanol for 30 minutes. The sections were then blocked in 3% horse serum for 20 minutes and incubated with a primary antibody, ab3778, for 1 hour at a 1/50 dilution. After primary antibody incubation the sections were incubated with a streptavidin linked horse anti-mouse IgG secondary antibody for 30 minutes. After secondary antibody incubation, the sections were incubated with avidin-biotinylated enzyme complex for 30 minutes, and then VIP substrate (purple color) was applied on sections for 4 minutes.

    The image shown is at 4 weeks, 25M.

References

This product has been referenced in:
  • Vega SL  et al. Combinatorial hydrogels with biochemical gradients for screening 3D cellular microenvironments. Nat Commun 9:614 (2018). ICC/IF . Read more (PubMed: 29426836) »
  • Yeung P  et al. A human osteoarthritis osteochondral organ culture model for cartilage tissue engineering. Biomaterials 162:1-21 (2018). IHC-P . Read more (PubMed: 29428675) »
See all 38 Publications for this product

Customer reviews and Q&As

1-10 of 17 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.15% · Temperature: RT°C
Antigen retrieval step
Heat mediated
Sample
Rhesus monkey Tissue sections (Ovarian Section)
Specification
Ovarian Section
Permeabilization
No
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 22 2013

Question
Answer



Sorry but we don't have a standard ELISA procedure for use with this antibody. However I would recommend using an indirect ELISA approach. This is outlined here https://www.abcam.com/index.html?pageconfig=resource&rid=11389

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Answer

Thank you for contacting us.

Unfortunately the indicated primary antibodies have not been tested with Flow Cytometry, and therefore are not covered by our Abpromise guarantee.

I would instead recommend using a different antibody against Collagen II that has been tested in this application:

https://www.abcam.com/Collagen-II-antibody-ab116242.html

I am afraid none of the antibodies against Aggrecan in our catalogue have been tested in Flow Cytometry, but if you wish to test them and share your results with us (through the Abreview system) we will be happy to offer a discount code for a future purchase.

If you would like to participate in the Abreview system and test ab3778 with Flow Cytometry, please let me know before placing the order so I can raise the discount code for you.

To find out more about our Abreview system, please visit: https://www.abcam.com/abreviews

Ab97022 is a suitable secondary for recognising Mouse primaries with Flow Cytometry. The only recommendation for this particular antibody is using it at dilutions 1/50-1/200. For general Flow Cytometry protocols I would encourage you to have a look at our Website’s protocols which may be very helpful (open the scroll down menu under the Flow Cytometry / FACS tab to have access to all the FC protocols).

https://www.abcam.com/index.html?pageconfig=popular_protocols

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information

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Question
Answer

Vielen Dank für Ihren Anruf.



Leider habe ich eine schlechte Nachricht für Sie: Aggrecan hat insgesamt eine Übereinstimmung von 19%, es gibt eine Region zwischen AS 800-850 des menschlichen Proteins, welches eine hohe Übereinstimmung mit dem Ziegen Protein hat, nur haben wir leider keinen passenden Antikörper.

Versican und SOX5 weisen leider gar keine Übereinstimmung mit dem Ziegenprotein auf.

Da alle unsere Aggrecan,Versican und SOX5Antikörper gegen das humane Protein entwickelt worden sind, kannich Ihnen leider keinen Vorschlag machen.

Ich hoffe, dies hilft Ihnen trotzdem weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

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Answer

Thank you for your inquiry.


We have quite a few references for this antibody to be used in ICC/IF and IHC-Fr/IHC-P. You can find them listed here and they are linked to the PubMed IDs.


https://www.abcam.com/Aggrecan-antibody-6-B-4-ab3778-references.html


You may also be interested in reading our customer reviews, called Abreviews, which include their protocol and images.


https://www.abcam.com/Aggrecan-antibody-6-B-4-ab3778-reviews.html


I hope this information helps. Please contact us with any other questions.

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Answer

Gracias por contactarnos.

Varios marcadores pueden usarse para la detección de condrocitos mediante immunohistoquímica. Solo algunos de ellos se han testado en especie canina, el resto me temo que no se han testado, por lo que se desconoce su funcionamiento en ellas.

Le indico algunas posibilidades con los números de catálogo de Abcam:

Collagen II: podría usarse ab300 o ab34712. Ninguno de ellos se ha testado en perro. Si está interesado en usarlo en esta especie, quizás le interese participar en el sistema Abreviews de evaluación de anticuerpos, por el cual les ofrecemos un anticuerpo completamente gratuito al mandarnos los resultados de testar el anticuerpo en la nueva especie. Si le interesa, por favor, comuníquemelo antes de realizar el pedido.

Para saber más sobre el sistema Abreviews: https://www.abcam.com/abreviews

Aggrecan: ab3778. Tampoco testado en perro, pero susceptible de evaluar su aplicación.

CD29: ab30388. En este caso tampoco hay datos de su uso en especie canina, tendría que evaluarlo usted misma.

CD44: ab19622 y ab119863. Ninguno de ellos se ha testado en perro, sin embargo, debido al elevado porcentaje de homología con dicha especie, es probable que reaccione. Por ello, si le interesa comprobarlo, puede igualmente usarlo y enviarnos los resultados, beneficiándose de una futura compra totalmente gratuita.

SOX9: ab26414 y ab71762. En este caso, ambos anticuerpos se han usado y caracterizado en especie canina, por lo que están cubiertos por la garantía Abpromise.

Espero que esta información le sea de utilidad. Si necesita que le enviemos una proforma de alguno o varios de los anticuerpos, si necesita más información de alguno de ellos, o si quiere saber más acerca del sistema Abreview, por favor, no dude en volvernos a contactar.

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Answer

Thank you for contacting us.

For the aggrecan antibody ab3778, I recommend the following two papers, if they are avalable to you, the first of which describes production of this clone and its use for IHC. We do not have access to the journal.

Caterson B et al. Mechanisms involved in cartilage proteoglycan catabolism. Matrix Biol 19:333-44 (2000). PubMed: 10963994

Little CB et al. Matrix metalloproteinases are involved in C-terminal and interglobular domain processing of cartilage aggrecan in late stage cartilage degradation. Matrix Biol 21:271-88 (2002). PubMed: 12009333

Please also note the review of ab3778 submitted by a customer on March 21, 2007, who included this note:

"We tried multiple retreival methods and found that pronase and hyaluronidase will work. The best results were with hyaluronidase".

(I suggest trying the hyaluronidase solution that is mentioned below for use with the collagen II antibody: 2mg/ml hyaluronidase in TBS for 2 hours at 37C).

For the collagen II antibody, an Abcam customer submitted an Abreview with the following protocol for IHC of formalin-fixed paraffin-embedded human cartilage tissue:

Antigen retrieval step: Enzymatic - Buffer/Enzyme Used: 2mg/ml hyaluronidase with 0.125U/ml chondroitinase in TBS for 2hrs 37C.

Blocking step: Normal goat serum in 1% BSA as blocking agent for 30 mins · Concentration: 20 % Temperature: 21°C.

Dilution of antibody: 1/50

Incubation time: 18 hours

Temperature: 4°C

Diluent: Blocking solution.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you very much for your email. Indeed, I have to agree with you that there is a problem. I would like to apologize for this and would like to reassure you that we will investigate this further and take the appropriate actions. The mouse reactivity has been added due to a customer feedback. Doing however the alignment of the immunogen with the mouse sequence, there is little overlap and it looks therefore rather tricky to get a specific staining on mouse cells. ** *.:*:***: http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111207-093227-0400-33224393-pg I will remove the mouse reactivity and replace this antibody for you with another antibody of your choice. Please do provide me therefore with the order details, such as the order number or the order date and name of the person placing the order. Unfortunately, we do have currently however no anti aggrecan antibody tested in your application and species. If you would like nevertheless to use one of our other anti aggrecan antibodies, I would be pleased to make predictions for the suitability. I apologize again for this error and am looking forward to hear back from you.

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Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (MEF, R1 ES cells, E14 ES cells)
Specification
MEF, R1 ES cells, E14 ES cells
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton

Abcam user community

Verified customer

Submitted Jun 20 2011

Answer

Thank you for your enquiry. Unfortunately, we do not have a Certificate of Analysis for this product. If there is anything else that I can help you with, please do not hesitate to contact me.

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1-10 of 17 Abreviews or Q&A

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