製品の概要

  • 製品名
    Anti-CD68 antibody [KP1]
    CD68 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [KP1] to CD68
  • 由来種
    Mouse
  • アプリケーション
    適用あり: ICC/IF, IHC-FoFr, IHC-P, IHC-Frmore details
  • 種交差性
    交差種: Human
    交差が予測される動物種: Mouse, Rat, Rabbit
  • ポジティブ・コントロール
    • IHC-P: Human tonsil tissue.
  • 特記事項

    We have received mixed customer feedback regarding suitability of ab955 with mouse and rat tissue and therefore no longer recommend it for mouse and rat. Please note it will also not be covered by our AbPromise guarantee for these species.

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファー
    Preservative: 0.099% Sodium azide
    Constituents: 0.9% Proprietary component, Water
  • Concentration information loading...
  • 精製度
    Protein A/G purified
  • ポリ/モノ
    モノクローナル
  • クローン名
    KP1
  • ミエローマ
    unknown
  • アイソタイプ
    IgG1
  • 軽鎖の種類
    kappa
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab955 in the following tested applications.

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

アプリケーション Abreviews 特記事項
ICC/IF Use at an assay dependent concentration. PubMed: 18804859
IHC-FoFr Use at an assay dependent concentration.
IHC-P 1/100 - 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Recommended heat mediated antigen retrieval in pressure cooker for 2-5 minutes or steaming of tissue sections for 45-60 minutes followed by cooling for 10 minutes and washing in distilled water.

In case of enzymatic epitope retrieval - digest with Pepsin enzyme for 30-60 seconds at RT.

IHC-Fr Use at an assay dependent concentration.

ターゲット情報

  • 機能
    Could play a role in phagocytic activities of tissue macrophages, both in intracellular lysosomal metabolism and extracellular cell-cell and cell-pathogen interactions. Binds to tissue- and organ-specific lectins or selectins, allowing homing of macrophage subsets to particular sites. Rapid recirculation of CD68 from endosomes and lysosomes to the plasma membrane may allow macrophages to crawl over selectin-bearing substrates or other cells.
  • 組織特異性
    Highly expressed by blood monocytes and tissue macrophages. Also expressed in lymphocytes, fibroblasts and endothelial cells. Expressed in many tumor cell lines which could allow them to attach to selectins on vascular endothelium, facilitating their dissemination to secondary sites.
  • 配列類似性
    Belongs to the LAMP family.
  • 翻訳後修飾
    N- and O-glycosylated.
  • 細胞内局在
    Cell membrane and Endosome membrane. Lysosome membrane.
  • Information by UniProt
  • 参照データベース
  • 別名
    • CD 68 antibody
    • CD68 antibody
    • CD68 antigen antibody
    • CD68 molecule antibody
    • CD68_HUMAN antibody
    • DKFZp686M18236 antibody
    • gp11 antibody
    • Gp110 antibody
    • LAMP4 antibody
    • Macrophage antigen CD68 (microsialin) antibody
    • MACROPHAGE ANTIGEN CD68 antibody
    • Macrosialin antibody
    • SCARD1 antibody
    • Scavenger receptor class D member 1 antibody
    see all

画像

  • ab955 staining CD68 in human spleen tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

    Tissue was fixed with formaldehyde and blocked with 10% serum for 10 minutes at 20°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/200) for 30 minutes at 20°C. An undiluted HRP-conjugated rat anti-mouse/rabbit polymer was used as the secondary antibody.

    See Abreview

  • ab955 staining CD68 in human liver tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

    Tissue was fixed with paraformaldehyde. Samples were then blocked with 10% serum for 3 hours at 22°C followed by incubation with the primary antibody at a 1/100 dilution for 16 hours at 4°C. An Alexa-Fluor® 568 conjugated goat anti-mouse polyclonal was used as secondary antibody at a 1/400 dilution.

    See Abreview

  • ab955 staining CD68 in human ulcerated oral (Mucosa/Bone) tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

    Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Triton-X 100 in PBS and blocked with 2.5% serum for 90 minutes at 25°C; antigen retrieval was by heat mediation in citrate buffer (pH 6). Samples were incubated with primary antibody (1/500 in 1% serum in PBS +0.01% Triton-X 100) for 16 hours at 4°C. A commercial IHC kit and DAB was used to visualize the staining.

    See Abreview

  • ab955 staining CD68 in human lung cancer tissues by ICC/IF (Immunocytochemistry/immunofluorescence).

    Cells were fixed with formalin and blocked with 2% horse serum for 1 hour at room temperature. Samples were then incubated with primary antibody (1/100) for 1 hour at 21°C. An Alexa Fluor® 488-conjugated Donkey anti-mouse IgG polyclonal (1/500) was used as the secondary antibody. DAPI containing mounting medium was used for nuclear staining (blue) and anti-TREM-1 (1/500) used for TREM-1 staining (red).

  • Immunohistochemical analysis of tonsil labeling CD68 with ab955.

  • Immunohistochemical analysis of tonsil labeling CD68 with ab955.

参考文献

This product has been referenced in:
  • You S  et al. An Aza resveratrol-chalcone derivative 6b protects mice against diabetic cardiomyopathy by alleviating inflammation and oxidative stress. J Cell Mol Med 22:1931-1943 (2018). Read more (PubMed: 29327811) »
  • Kim SJ  et al. Anti-inflammatory effect of Tauroursodeoxycholic acid in RAW 264.7 macrophages, Bone marrow-derived macrophages, BV2 microglial cells, and spinal cord injury. Sci Rep 8:3176 (2018). Read more (PubMed: 29453346) »
See all 169 Publications for this product

レビューと Q&A

1-10 of 16 Q&A

Answer

The IgG concentration of ab955, Anti-CD68 antibody [KP1], lot GR173584-2 is 4µg/mL.

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Answer



Any products that require an MSDS form will have a link on the product's online datasheet to the data. It will be a PDF document link labeled 'SDS'. Please see attached image for example of what to look for on the datasheet. If no 'SDS' link is available than the product does not contain any components that require an SDS.

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Question
Answer

Thank you very much for your calls this week and for letting us know about the trouble with this antibody in IHC-Fr.

As we discussed, I'm sending a free of charge replacement vial of ab125212 on the order ***. This should arrive on Monday.

Please keep me updated about the results with this new antibody, and let me know if you have any questions or if there is anything else that we can do for you.

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Question
Answer

Thank you for bringing this to our attention. Please contact me if the cold acetone (on ice, 10 minutes) fixation that I recommended does not improve your result.

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Answer

Thank you for your enquiry.

I can confirm that the immunogen used for CD68 antibody [KP1] (ab955) is lysosomal fraction of human lung macrophages.

I am sorry this has not been specifically epitope mapped and we have no further information on this occasion.

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

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Answer

Muchas gracias por tu respuesta.

Es importante que todos los anticuerpos que escojas para llevar a cabo IHC-P en tejido humano estén validados (y por tanto garantizados por Abcam) para funcionar en esta especie y aplicación. Esta información aparece en la hoja técnica de los productos.

También es importante echar un vistazo a las hojas de datos de cada uno de los productos para tener en cuenta recomendaciones especificas, tales como diluciones, métodos de recuperación antigénica, y demás que pueden aparecer en ellas. A las fichas técnicas se puede acceder desde la página web, escribiendo el código del anticuerpo (ab955, por ejemplo) en la casilla azul en la parte de arriba de nuestra página web (https://www.abcam.com/).

Desconozco las combinaciones que queréis llevar a cabo mediante inmunofluorescencia doble con los anticuerpos mencionados. En general, a la hora de hacer inmunoensayos múltiples hay que tener mucha precaución para evitar reactividad cruzada. Es importante usar anticuerpos procedentes de distintas especies si se puede, o al menos con diferentes isotipos. Una buena forma de evitar reactividad cruzada es usando secundarios de la misma especie. Además esto permite usar suero de dicha especie para bloquear.

Te copio los links a los protocolos que tenemos en nuestra web para llevar a cabo doble inmunofluorescencia por s pudieran resultar de ayuda:

https://www.abcam.com/index.html?pageconfig=resource&rid=11459

https://www.abcam.com/index.html?pageconfig=resource&rid=11458

De todas maneras si quieres que comentemos distintas posibilidades y escenarios posibles, no dudes en contactarme otra vez.

Respecto a los secundarios, tenemos muchos anticuerpos que podrían serviros. Te aconsejo que en función de los primarios y combinaciones que vayáis a llevar a cabo, optéis por uno u otro. Para llevar a cabo la búsqueda de los secundarios desde la pagina web (https://www.abcam.com/) pincha en la pestaña “Secondary Antibodies”. Se abre una nueva ventana de “Advance Search”, en la cual se pueden elegir las categorías requeridas. La única categoría obligatoria es el isotipo del anticuerpo, pues tiene que coincidir con el del primario. El resto de categorías son opcionales, pero son muy útiles para afinar la búsqueda y elegir el anticuerpo más apropiado según la especie del primario, la conjugación deseada, la clonalidad, la aplicación en la que se va a testar, etc…

Los precios y la disponibilidad pueden consultarse igualmente en la parte superior de la datasheet de los productos. Si necesitas un presupuesto no dudes en hacérmelo saber para que te lo envíe. Además tenemos algún descuento para pedidos superiores a 5 productos, por lo que si vais a realizar un pedido de estas características te animaría a que nos contactaras primero.

Espero que esta información sea de ayuda. Como te digo, para cualquier aclaración o más sugerencias, estaré encantada de echar una mano.

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Answer

Thank you for contacting us.

Effectively all of the products mentioned are suitable, and therefore are guaranteed for IHC in frozen sections in the species stated on their datasheets.

However, instead of using PFA as a fixative, I would better suggest using acetone, methanol or ethanol at -20°C for 30 min, so the additional antigen retrieval step can be skipped. If, by the tissue’s nature PFA is required as a fixative, antigen retrieval is recommended to allow the antibody reach the epitope.

All of our products are covered by our Abpromise® guarantee, which ensures that you can trust them, and they should work in the tested species and applications stated on the datasheet, or we will offer a replacement, credit, or refund, if reported within 6 months of purchase.

To read about our Abpromise policy: https://www.abcam.com/index.html?pageconfig=abpromise

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 for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab955 since this antibody has been tested in mouse paraffin-embedded tissue sections.


For the fixation, we generally recommend between 18 - 24 hrs. It may be just that your sections are underfixed.


For antigen retrieval, was this performed in the microwave? We generally recommend 20 mins, but it's best to optimize between 15 - 30 mins. When the 20 mins has elapsed, you can run cold tap water into the vessel for 10 mins.


If you want to permeablize the sections, you should do a 0.1% Triton-X 100 in PBS treatment as a separate step and omit the Triton X-100 from the blocking buffer.


For blocking, have you tried5-10% normal goat serum instead of BSA? You should also put this in PBS.


The most important thing to try wouldbe to test a1/100 dilution of ab955 overnight at 4C instead of 1/200 for 1 hr RT. You should do this incubation in 1% BSA in PBS.


For the s100 antibody ab8330, would you be able to run a positive control with rat tissues since this antibody isn't tested in mouse?


For the CD31 antibody, ab7388, we know that this does not work well with PFA fixation (see Abreview linkedbelow). We have heard that it works wellwhen used on frozen sections of mouse skin that werefixed in methanol:acetone for 10 mins. Would you be able to try some frozen methanol or acetone fixed sections as a positive control since IHC-P does not work.


https://www.abcam.com/index.html?datasheet=7388&tab=abreviews&intabreviewid=23226

Should the suggestions not improve the results, please do let me know.

In the event that a ab955 is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund for this tested antibody.

I hope this information is helpful, and I thank you for your cooperation.

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Answer

Thank you very much for your interest in anti-CD68 antibody clone [KP1] (ab955).

To our knowledge, ab955 has not been tested in Western blot, but I do not see a reason why it should not work. Therefore, I can offer a discount off a future purchase if you buy ab955 now, test it in Western blot and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab955 in Western blot. I will then send a discount code. This code must be issued before purchasing ab955 so please wait for my reply before ordering.

2. Purchase ab955 either by phone, fax, or online (www.abcam.com).

3. Test it in Western blot.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out more about our Abreview system, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any primary antibody ordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab955 turns out to be unsuitable for Western blot, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount

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Answer

Thank you for contacting Abcam.

I feel that both products are suitable for IHC-P. Both have many specific references and in-depth scientific support data right on the datasheet. They differ mainly in the species in which they were raised and in the tags used for detection. Ab6728 is an HRP conjugate whereas Ab6788 is Biotin labeled. I believe that you should choose the secondary which has an associated detection system that you are most familiar with in case you should run into any staining issues.

I hope this information is helpful. Please let me know if you have any questions.

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1-10 of 16 Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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