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Recombinant

Recombinant Anti-Aggrecan antibody [6-B-4] (ab3778)

Reviews (6)Q&A (12)References (129)
Western blot - Anti-Aggrecan antibody [6-B-4] (ab3778)
  • Western blot - Anti-Aggrecan antibody [6-B-4] (ab3778)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aggrecan antibody [6-B-4] (ab3778)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aggrecan antibody [6-B-4] (ab3778)
  • Anti-Aggrecan antibody [6-B-4] (ab3778)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Mouse monoclonal [6-B-4] to Aggrecan
  • Suitable for: WB, IHC-P
  • Reacts with: Human, Recombinant fragment

Conjugates logo Related conjugates and formulations

Carrier Free

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

  • Tested applications

    Suitable for: WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Human, Recombinant fragment

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Recombinant human Aggrecan protein. C-terminal 10 His tagged Human Aggrecan recombinant protein (aa 20 to 675). IHC-P: Human cartilage sections.

  • General notes

    This product has switched from a hybridoma to recombinant production method on 08th October 2020.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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 long term. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59.04% PBS
  • Concentration information loading...

  • Purity

    Protein A 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

  • Isotype

    IgG1

  • Light chain type

    kappa

  • Research areas

Applications

The Abpromise guarantee

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
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 250 kDa.
IHC-P (3)
Use a concentration of 0.05 µg/ml. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.
Notes
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 250 kDa.
IHC-P
Use a concentration of 0.05 µg/ml. Perform enzymatic antigen retrieval before commencing with IHC staining protocol.

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

    • Western blot - Anti-Aggrecan antibody [6-B-4] (ab3778)
      Western blot - Anti-Aggrecan antibody [6-B-4] (ab3778)
      Anti-Aggrecan antibody [6-B-4] (ab3778) at 1/1000 dilution + C-terminal 10 His tagged Human Aggrecan recombinant protein (aa 20 to 675) 15ng

      Secondary
      Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

      Predicted band size: 250 kDa



      Buffer and Concentration: 2% BSA/TBST

      Exposure time: 180 seconds

    • Western blot - Anti-Aggrecan antibody [6-B-4] (ab3778)
      Western blot - Anti-Aggrecan antibody [6-B-4] (ab3778)

      ab3778 was shown to react with recombinant human Aggrecan protein in western blot. 

      Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab3778 at 1µg/ml overnight at 4°C. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye®800CW) preabsorbed (ab216772) at a 1/20000 dilution for 1 hour at RT.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aggrecan antibody [6-B-4] (ab3778)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aggrecan antibody [6-B-4] (ab3778)

      Secreted staining of cartilage cells.

      Staining performed on a Leica BOND RXTM system using the standard protocol F. The section was pre-treated using enzymatic retrieval, for 15mins. The section was then incubated with ab3778, 0.05µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aggrecan antibody [6-B-4] (ab3778)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aggrecan antibody [6-B-4] (ab3778)

      No staining in negative control tissue (human normal liver).

      Staining performed on a Leica BOND RXTM system using the standard protocol F. The section was pre-treated using enzymatic retrieval, for 15mins. The section was then incubated with ab3778, 0.05µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    • Anti-Aggrecan antibody [6-B-4] (ab3778)
      Anti-Aggrecan antibody [6-B-4] (ab3778)

    References (129)

    Publishing research using ab3778? Please let us know so that we can cite the reference in this datasheet.

    ab3778 has been referenced in 129 publications.

    • Zhao K  et al. Acid-sensing ion channels regulate nucleus pulposus cell inflammation and pyroptosis via the NLRP3 inflammasome in intervertebral disc degeneration. Cell Prolif 54:e12941 (2021). PubMed: 33111436
    • Lu X  et al. miR-335-5P contributes to human osteoarthritis by targeting HBP1. Exp Ther Med 21:109 (2021). PubMed: 33335572
    • Guo Q  et al. STING promotes senescence, apoptosis, and extracellular matrix degradation in osteoarthritis via the NF-?B signaling pathway. Cell Death Dis 12:13 (2021). PubMed: 33414452
    • Li C  et al. LncRNA NEAT1 promotes nucleus pulposus cell matrix degradation through regulating Nrf2/ARE axis. Eur J Med Res 26:11 (2021). PubMed: 33478594
    • Wang W  et al. Positive feedback regulation between USP15 and ERK2 inhibits osteoarthritis progression through TGF-ß/SMAD2 signaling. Arthritis Res Ther 23:84 (2021). PubMed: 33726807
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a question

    1-10 of 12 Q&A

    Question

    send protocol for ELISA

    Read More
    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

    Read More

    Question

    Dear Sir,
    The customer will order ab3778, ab3092, ab97022 for Flow cytometry application, sample is human PBMC. So,she would like to abcam guide for Flow cytometry method ( staining step?, wash step? )

    Read More
    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

    Read More

    Question

    Welcher Antikoerper ist geignet fuer Ziege und Human, IHC-P: Versican, SOX5, Aggrecan?

    Read More
    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.

    Read More

    Question

    I want to use this antibody for immunofluorescenceon my human cell samples: i) monolayer on coverslip and ii) frozen sections, both previously fixed with 2% PFA. Is there any literature that have the same application using this antibody? I would like to take reference from the protocol they used, including the details like the antibody dilution ratio and the antigen retrieval steps). I would be grateful if you can refer to me the NCBI code of these papers and suggest a suitable protocol. Thank you very much.

    Read More
    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.

    Read More

    Question

    Por favor, indiquenos unos anticuerpos validos para la siguiente situacion:
    Applications: Immunochemestry
    Reacts with: Chondrocytes or precursors of chondrocytes
    Sample: canine (dog) trachea
    Muchas gracias

    Read More
    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.

    Read More

    Question

    Please send the antigen retrieval protocol for the aggecan antibody ab3778 and the collagen II antibody ab34712.

    Read More
    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.

    Read More

    Question

    LOT NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM No staining SAMPLE Mouse cartilage PRIMARY ANTIBODY Dilution 1/50 in PBS/BSA 1% for 1 hour at RT Wash with PBS DETECTION METHOD DAB ANTIBODY STORAGE CONDITIONS Short term at 4°C Long term at -20°C or -80°C FIXATION OF SAMPLE Paraformaldehyde 24hours ANTIGEN RETRIEVAL Enzymatic method: Hyaluronidase PERMEABILIZATION STEP no BLOCKING CONDITIONS BSA 1% 30min at 37°C SECONDARY ANTIBODY Vectastain ABC kit HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes ADDITIONAL NOTES In the scientific support, in your answer at the question 4 you say that these Ab is only react with human and cow! But in the data sheet you say that it react with mouse???

    Read More
    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.

    Read More

    Question

    Do you have the certificate of analysis for Lot＃：196340、Lot＃：346985、Lot＃：393795?

    Read More
    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.

    Read More

    Question

    Which antibody is most suitable for detecting full length aggregan protein in goat tissue?

    Read More
    Answer

    I have been informed of the following information which I hope will help you: Ab3773: this should be reactive with goat since this antibody recognises a sequence ARG... that is conserved in all species Guinea Pig/rat to human but not mouse for some reason where the sequence is ALG.... This antibody will not recognise the sequence ...ARG.. if it is in the non-cleaved intact aggrecan protein core; i.e. it will only recognise the aggrecanase generated neoepitope ARG... Ab3778: this antibody recognises a linear sequence ..EPEEP... that is present in the IGD protein core of human and bovine aggrecan but not pig and several other animals; it seems it is not present in goat protein after BLASTing the sequence in goat. This antibody recognises the linear epitope ...EPEEP... in both intact and matrix protease degraded aggrecan from human and bovine cartilage. Please do not hesitate to contact me if I can be of further help,

    Read More

    Question

    In native human cartilage tissue, whether it stains the extracellular region, or pericellular region or intracellular region?

    Read More
    Answer

    Thank you for your enquiry regarding ab3778. I'm afraid we do not have an image of the staining of human cartilage with this antibody. However, there are two papers published with this product which hopefully will have some information to answer your question. 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 Caterson B et al. Mechanisms involved in cartilage proteoglycan catabolism. Matrix Biol 19:333-44 (2000). PubMed: 10963994 Unfortunately I cannot access those to find it out myself, I apologise for the inconvenience. Two reviews of this antibody used on human cartilage were also published on the datasheet, and you can contact one of the scientists via this method via the online datasheet, I hope Lynsey Fairbairn can also provide information on the staining she observed. I hope this information will help you, I'm sorry I couldn't help you more on this occasion,

    Read More

    1-10 of 12 Q&A

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