Anti-CCR4抗体(ab1669)

Reviews (4)Q&A (7)References (12)

Key features and details

  • Goat polyclonal to CCR4
  • Suitable for: WB
  • Reacts with: Human, Cynomolgus monkey
  • Isotype: IgG

概述

  • 产品名称

    Anti-CCR4抗体
    参阅全部 CCR4 一抗

  • 描述

    山羊多克隆抗体to CCR4

  • 宿主

    Goat

  • 特异性

    Peptide sequence is < 50 % identical to other human chemokine receptors in this region.

  • 经测试应用

    适用于: WBmore details

  • 种属反应性

    与反应: Human, Cynomolgus monkey

  • 免疫原

    Synthetic peptide:

    SNYYLYESIPKPCTKEGIKAFGE

    , corresponding to amino acids 17-39 of Human CCR4 (extracellular domain).

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • 常规说明

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

性能

应用

Our Abpromise guarantee covers the use of ab1669 in the following tested applications.

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

应用 Ab评论 说明
WB 1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 42 kDa).

靶标

  • 功能

    High affinity receptor for the C-C type chemokines CCL17/TARC and CCL22/MDC. The activity of this receptor is mediated by G(i) proteins which activate a phosphatidylinositol-calcium second messenger system. Can function as a chemoattractant homing receptor on circulating memory lymphocytes and as a coreceptor for some primary HIV-2 isolates. In the CNS, could mediate hippocampal-neuron survival.

  • 组织特异性

    Predominantly expressed in the thymus, in peripheral blood leukocytes, including T-cells, mostly CD4+ cells, and basophils, and in platelets; at lower levels, in the spleen and in monocytes. Detected also in macrophages, IL-2-activated natural killer cells and skin-homing memory T-cells, mostly the ones expressing the cutaneous lymphocyte antigen (CLA). Expressed in brain microvascular and coronary artery endothelial cells.

  • 序列相似性

    Belongs to the G-protein coupled receptor 1 family.

  • 翻译后修饰

    In natural killer cells, CCL22 binding induces phosphorylation on yet undefined Ser/Thr residues, most probably by beta-adrenergic receptor kinases 1 and 2.

  • 细胞定位

    Cell membrane.
  • Information by UniProt

  • 数据库链接

    • 别名

      • C C chemokine receptor type 4 antibody
      • C C CKR 4 antibody
      • C-C chemokine receptor type 4 antibody
      • C-C CKR-4 antibody
      • CC CKR 4 antibody
      • CC-CKR-4 antibody
      • CCR 4 antibody
      • CCR-4 antibody
      • CCR4 antibody
      • CCR4_HUMAN antibody
      • CD194 antibody
      • Chemokine (CC motif) receptor 4 antibody
      • chemokine C C motif receptor 4 antibody
      • ChemR13 antibody
      • CKR4 antibody
      • CMKBR 4 antibody
      • CMKBR4 antibody
      • HGCN 14099 antibody
      • K5 5 antibody
      • K5-5 antibody
      • MGC88293 antibody
      see all

    图片

    • Western blot - Anti-CCR4 antibody (ab1669)
      Western blot - Anti-CCR4 antibody (ab1669)
      Anti-CCR4 antibody (ab1669) at 1 µg/ml + Human thymus tissue lysate - total protein (ab30146) at 10 µg

      Secondary
      Rabbit polyclonal to Goat IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 42 kDa
      Observed band size: 55 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 100 kDa, 37 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 12 minutes


      CCR4 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

    文献 (12)

    发表研究结果有使用 ab1669？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab1669 被引用在 12 文献中.

    • Yuan FY  et al. Tanshinone IIA improves diabetes mellitus via the NF-?B-induced AMPK signal pathway. Exp Ther Med 16:4225-4231 (2018). PubMed: 30344697
    • Wang Y  et al. HIC1 deletion promotes breast cancer progression by activating tumor cell/fibroblast crosstalk. J Clin Invest 128:5235-5250 (2018). PubMed: 30204129
    • Berlato C  et al. A CCR4 antagonist reverses the tumor-promoting microenvironment of renal cancer. J Clin Invest 127:801-813 (2017). PubMed: 28134623
    • Maolake A  et al. Tumor-associated macrophages promote prostate cancer migration through activation of the CCL22-CCR4 axis. Oncotarget 8:9739-9751 (2017). Human . PubMed: 28039457
    • Ou B  et al. CCR4 promotes metastasis via ERK/NF-?B/MMP13 pathway and acts downstream of TNF-a in colorectal cancer. Oncotarget 7:47637-47649 (2016). IHC . PubMed: 27356745
    View all Publications for this product

    客户评价及客户问答

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    提交评价 提交问题

    1-10 of 11 Abreviews or Q&A

    Question

    Bonjour,
    Merci de votre rapidité de réponse.
    Je vous retourne en échange votre document complété et mes images de blots.
    N'hésitez pas à me faire des remarques sur ceux ci pour augmenter mes chances de réussite avec le nouvel Ac.
    Dans la fiche technique de l'Ac que vous me faites parvenir (ab113085), le peptide ciblé (séquence, localisation sur la protéine) n'est pas précisé, le connaissez vous?
    Avez vous des publications référencées avec cet Ac car j'avais choisi l'ab1669 pour cela?
    Cordialement,

    Read More
    Answer

    Merci beaucoup pour toutes ces informations détailléesqui nous sont très utiles pour identifier l'origine du problème rencontré avec le ab1669 lot GR34771-3.

    Je n'ai rien à modifier à votre protocole qui me semble être optimal. La quantité de protéines (30µg) est ce que nous recommandons, le test de différentes dilutions d'anticorps primaire ainsi que l'utilisation de différents agents bloquants (lait ou BSA) sont également essentiels; parfait!

    Nous n'avons malheureusement pas encore trouvé de publication faisant référence à l'anticorps ab113085. Quant au peptide utilisé comme immunogène, je vais demander au laboratoire produisant cet anticorps si la séquence peut être communiquée.

    Je vous contacterai de nouveau dès que j'aurai reçu une réponse.

    Merci pour votre patience et n'hésitez pas à m'informer des résultats obtenus avec l'anticorps de remplacement ab113085.

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    Question


    Nous vous avons acheté un Ac fin avril et après de multiples essais celui ci ne
    révèle pas la protéine souhaitée (+ beaucoup de non spécifique):
    Anticorps anti-CCR4 (ab1669)
    Cellules testées = cellules humaines
    Application WB
    Pouvez vous me dire qu'elles sont les modalités / conditions pour remplacer cet
    Ac par un autre?
    Je serais éventuellement intéressée pour tester soit l'ab83250 ou l'ab113085
    selon vos conseils.
    Cordialement,

    Read More
    Answer

    Merci de nous avoir contactés.

    Nous sommes désolés d'apprendre que le produit ab1669 que vous avez reçu ne fonctionne pas comme attendu.

    Nos produits sont garantis pour fonctionner tels qu'ils sont décrits sur la fiche technique, nous pouvons donc effectivement résoudre le problème que vous avez rencontré en vous envoyant un anticorps de remplacement.

    Les 2 anticorps ab83250et ab113085 (comme ab1669) sont garantis pour fonctionner en Western Blot et sur échantillons humains.
    L'anti-CCR4 ab83250 n'étant pas actuellement en stock, j'ai mis en place l'envoi d'une unité de ab113085. Le numéro de commande de remplacement gratuit de ce produit est *******. Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition.

    En échange, nous serions ravis si vous pouviez nous communiquer quelques détails concernant votre protocole en remplissant le questionnaire technique ci-joint. Ces informations sont très importantes pour nous car elles nous permettent d'investiguer la source du problème rencontré avec cet anticorps et de prendre les dispositions nécessaires pour assurer une bonne qualité de nos produits.

    N'hésitez pas à nous contacter lors d'une prochaine occasion.

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    Flow Cytometry abreview for Anti-CCR4 antibody

     Poor
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Cynomolgus Monkey Cell (Whole blood)
    Specification
    Whole blood
    Preparation
    Cell harvesting/tissue preparation method: After red blood cell lysis
    Sample buffer: PBS/BSA 1%/Azide 0.01%
    Fixation
    Paraformaldehyde
    Permeabilization
    No
    Gating Strategy
    alive lymphocytes
    Read More

    Abcam user community

    Verified customer

    提交于 Apr 27 2012

    Flow Cytometry abreview for Anti-CCR4 antibody

     Poor
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Rat Cell (Splenocytes)
    Specification
    Splenocytes
    Preparation
    Cell harvesting/tissue preparation method: spleen was mashed
    Sample buffer: PBS/BSA 1%/Azide 0.01%
    Permeabilization
    No
    Gating Strategy
    alive lymphocytes
    Read More

    Abcam user community

    Verified customer

    提交于 Apr 18 2012

    Flow Cytometry abreview for Anti-CCR4 antibody

     Good
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Mouse Cell (Whole blood)
    Specification
    Whole blood
    Preparation
    Cell harvesting/tissue preparation method: After red blood cell lysis
    Sample buffer: PBS/BSA 1%/Azide 0.01%
    Fixation
    Paraformaldehyde
    Permeabilization
    No
    Gating Strategy
    alive lymphocytes
    Read More

    Abcam user community

    Verified customer

    提交于 Apr 18 2012

    Flow Cytometry abreview for Anti-CCR4 antibody

     Average
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (PBMC)
    Specification
    PBMC
    Preparation
    Cell harvesting/tissue preparation method: after ficoll
    Sample buffer: PBS/BSA 1%/ Azide 0.01%
    Permeabilization
    No
    Gating Strategy
    on monocytes
    Read More

    Abcam user community

    Verified customer

    提交于 Dec 28 2011

    Question

    I have a customer who purchased your antibodies, ab1669 and ab1667. The customer would like to use them for immunohistochemistry of paraffin-embedded sections. Her sample is human lymphoid neoplasm. Please let me know an antigen retrieval method you recommend, respectively. And regarding each figure of IHC-formalin fixed, paraffin embedded sections please let me know also each antigen retrieval method when you got these data.

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    Answer

    Thank you for your enquiry. Ab1667: Antigen Retrieval: Sections were treated with antigen retrieval solution (Dako # S-1700, pH 6.0) and steamed for 20 minutes followed by cooling for 20 minutes. Variations of the method will work successfully as shown in the following publication that has images as well: http://clincancerres.aacrjournals.org/cgi/content/full/11/7/2459 Ab1669: Antigen Retrieval: Sections were treated with antigen retrieval solution (Dako # S-1700, pH 6.0 or #S-3308, pH 8.0) and steamed for 20 minutes followed by cooling for 20 minutes. I hope that this information helps. Should you have any further questions then please do not hesitate to get back in touch with me.

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    Question

    For each of these antibodies, specifically what type of antigen retrieval was used for IHC-formalin fixed, paraffin embedded sections?

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    Answer

    Thank you for your patience. The originator of ab12953 and ab13396 has gotten back to me with the standard IHC protocol that they follow which I have included below. If you have any additional questions, please let me know. Tissue Preparation: Formalin fixation and embedding in paraffin wax Tissue Sectioning: Make 4-µm sections and place on pre-cleaned and charged microscope slides. Heat in a tissue-drying oven for 45 minutes at 60°C. Deparaffinization: Wash dry slides in 3 changes of xylene – 5 minutes each @ RT Rehydration: Wash slides in 3 changes of 100% alcohol – 3 minutes each @ RT Wash slides in 2 changes of 95% alcohol – 3 minutes each @ RT Wash slides in 1 change of 80% alcohol – 3 minutes @ RT Rinse slides in gentle running distilled water – 5 minutes @ RT Antigen retrieval: Steam slides in 0.01 M sodium citrate buffer, pH 6.0 at 99-100°C - 20 minutes Remove from heat and let stand at room temperature in buffer - 20 minutes Rinse in 1X TBS with Tween (TBST) – 1 minute @ RT Immunostaining: (Do not allow tissues to dry at any time during the staining procedure) Apply a universal protein block – 20 minutes @ RT Drain protein block from slides, apply diluted primary antibody – 45 minutes @ RT Rinse slides in 1X TBST - 1 minute @ RT Apply a biotinylated anti-rabbit IgG (H+L) secondary – 30 minutes @ RT Rinse slides in 1X TBST - 1 minute @ RT Apply alkaline phosphatase streptavidin – 30 minutes @ RT Rinse slides in 1X TBST - 1 minute @ RT Apply alkaline phosphatase chromogen substrate – 30 minutes @ RT Wash slides in distilled water – 1 minute @ RT Dehydrate: (This method should only be used if the chromogen substrate is alcohol insoluble (e.g. Vector Red, DAB) Wash slides in 2 changes of 80% alcohol – 1 minute each @ RT Wash slides in 2 changes of 95% alcohol – 1 minute each @ RT Wash slides in 3 changes of 100% alcohol – 1 minute each @ RT Wash slides in 3 changes of xylene – 1 minute each @ RT Apply coverslip

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    Question

    For each of these antibodies, specifically what type of antigen retrieval was used for IHC-formalin fixed, paraffin embedded sections?

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    Answer

    Thank you for your phone call. For ab13268, ab1662, and ab1669, the originator has informed me that the antigen retrieval method for IHC-formalin fixed, paraffin embedded sections involved treating the sections with antigen retrieval solution (Dako # S-1700, pH 6.0 or #S-3308, pH 8.0) and steamed for 20 minutes followed by cooling for 20 minutes. I will contact you again when I receive information regarding ab12953 and ab13396. Have a great weekend!

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    Question

    I will try to answer your questions so you can help me. We have an answer: 1. Please describe the problem (high background, no staining etc). We found high background staining in all kryostat sections independent of the concentration of the primary antibody. 2. On what material are you testing the antibody in IHC? We made a positive control by injection of CCR4 positive CEM-cells (human leucaemia T-cell line) into human healthy skin. Then we made kryostat sections (6 µm). * Species? * Cell line? * Tissue? 3. How did you fix the samples? We fixed the samples before staining with acetone for 10 minutes. * Ethanol, methanol * Acetone * Paraformaldehyde * Other 4. Did you apply antigen retrieval step? We used a biotinylated secondary antibody, then ABC-kit (Vectastain). The substrate was DAB solution * Enzymatic method * Heat mediated technique * Other 5. How did you block the unspecific binding sites? We quenched endogenous peroxidases with 1% H2O2 (40 min). We blocked unspecific bindings with 1% bovine serum albumin (RT, humid chamber, 30 min). 6. Primary antibody The CCR4 antibody was a goat anti human CCR4 antibody. Dilutions: as descibed 1:300, we also tried 1:50, 1:150, 1:100, 1:500. We incubated the primary antibody overnight at 4° in a humid chamber. We washed in three steps with high volume (3 times a 10 min.) * Specification (in which species was it raised against)? * At what dilution(s) have you tested this antibody? * Incubation time, wash steps (multiple short washes are more effective than fewer longer wash steps)? 7. Secondary antibody We used a biotinylated anti-goat IgG antibody (produced in rabbit, Fa. Vector) Dilution 1:200, Incubation time 45 min. at RT, washing with PBS 2x10 min., 45 min. ABC reagent in humid chamber at RT, washing 2x10 min with PBS, 5 min. DAB solution. * What secondary antibody are you using? * Specification (in which species was it raised against)? * At what dilution(s) have you tested this antibody? * Incubation, wash steps? * Do you know whether the problems you are experiencing come from the secondary? * What detection method are you using? 8. Background staining See images of the attachment. "über" means overview. We here used the described antibody dilution 1:300. Only in this dilution we saw marked brown cells (CEM-cells). "pat1" means we stained skin from a patient with atopic dermatitis. * Please provide an image of your staining 9. Which detection system did you use? ABC-kit, Vaectastain 10. Did you apply positive and negative controls along with the samples? Please specify. yes, "patneg" means negative control (without primary antibody) Please see attachment. 11. Optimization attempts only a few times, we try to establish the protocol. We altered the washing steps (higher volumes of PBS), we tried different dilutions. We always saw high background staining. * How many times have you tried the IHC? * Do you obtain the same results every time? * What steps have you altered? We look forward to hearing from you soon.

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    Answer

    Thank you for submitting the completed questionnaire to us and for your patience. We are very sorry to hear that you are having problem with this antibody. We have searched our database and found that this is a popular selling product and your feedback is the first we have received about it not working. Therefore, at this stage, we would suggest that there is either a problem with the vial you received, or modifications to your protocol are needed to obtain a positive result. As we understand from your e-mails, you performed IHC on skin biopsies taken from patients with atopic dermatitis. We would strongly suggest using positive control along with the samples. Human spleen sections or human peripheral blood cells would be good for this purpose. This is especially important since there are some publications which suggest that expression of CCR4 in skin-derived lymphocytes is higher that its expression in other type of tissues. Lymphocytes from inflamed skin cells have especially high levels of CCR4 which may be one of the reasons why you are having such a high background signal. Please take a look at this published paper; you may find some useful information: http://ajp.amjpathol.org/cgi/content/full/160/1/347 You may also need to alter certain steps in your protocol. Try to increase the concentration of the blocking serum i.e. 3-5% BSA to see if the background signal is getting lower. Block the endogenous peroxidase activity with 1.6% (not 1%) H2O2, it may help. We have looked at the images you have forwarded to us, it is clear that the antibody works. Comparing the intensity of the signal in the negative control and in the samples, the staining is definitely stronger in the biopsy samples. Although, it is true that the background signal in some areas of the sections is a bit high. We hope this will be useful for you. Should you still have any problem, then please do not hesitate to contact us again.

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    1-10 of 11 Abreviews or Q&A

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