アブカムでは最適な動作のために Google Chrome など最新ブラウザでの閲覧を推奨します。

私たちはウェブサイトをできるだけ使いやすくするために、クッキーを使用しています。

クッキー・ポリシーでは、使用するクッキーをオプトアウトする方法について説明しています。

クッキーの設定を変更しないままでいる場合、このポリシーに同意しているとみなされます。

続ける 続ける

お問い合わせは電話 +81-(0)3-6231-0940 または メールでどうぞ

  • アカウント
  • ログアウト
ログイン または 新規登録

Welcome

ログイン または

すでにアカウントをお持ちですか?

新規登録
カート
クイック・オーダー
Abcam homepage

  • 研究用製品
    製品タイプ別
    一次抗体
    二次抗体
    イムノアッセイキット・試薬
    細胞・組織イメージング
    細胞・バイオケミカルアッセイ
    タンパク質・ペプチド
    製品タイプ別
    プロテオミクス
    アゴニスト・アクチベータ-・アンタゴニスト・インヒビター
    細胞株・ライゼート
    miRNA マルチプレックス・アッセイキット
    マルチプレックス・アッセイ
    研究分野別
    癌
    循環器系
    細胞生物学
    エピジェネティクス
    メタボリズム
    発生生物学
    研究分野別
    免疫学
    微生物学
    脳神経科学
    シグナル伝達
    ステムセル
  • 診断と治療のソリューション
    カスタム・ソリューション & パートナーシップ

    あなたの診断法や治療法を発展させるための、カスタム抗体開発およびコマーシャル・パートナーシップ

    私たちと共にカスタム・ソリューションを創造する

    私たちとパートナーになる

  • サポート
    サポートハブ

    研究のためのサポートとアドバイス

    サポートハブを表示

    プロトコール

    実験のステップごとの詳細

    プロトコールを表示

  • イベント
    • イベント・カレンダー
    • 癌
    • 循環器
    • エピジェネティクス&核内情報
    • 免疫学
    • ニューロサイエンス
    • ステムセル
    • 展示会
    • ウェビナー
    最新のイベント情報

    世界中でアブカムが主催する研究会やセミナーの日程、内容、演者など

    イベント・カレンダー

  • パスウェイ
    細胞シグナル伝達パスウェイ

    すべてのパスウェイを見る

    インタラクティブ・パスウェイを見る

  • ブログ
    アブカム・ブログ

    最新の情報はこちら.

    • キャンペーン
    • 製品情報
    • 技術情報

新型コロナウイルス (COVID-19)に伴う当社の対応について

  1. Link

    ccr7-antibody-ab1657.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Immunology Innate Immunity Chemokines Beta Chemokine Rec. (CCR)
Share by email

Anti-CCR7 抗体 (ab1657)

  • Datasheet
  • SDS
Reviews (4)Q&A (11)References (9)

Product price, shipping and contact information

Currently unavailable

申し訳ございませんが、現在表示できません。

こちらにお問い合わせいただくか、またはしばらく経ってから改めてお試しください

 

Loading size & price…

 

出荷および受注について

Abpromise

保証された製品品質、優れたカスタマー・サポート。 詳細はこちら。

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR7 antibody (ab1657)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR7 antibody (ab1657)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR7 antibody (ab1657)

Key features and details

  • Goat polyclonal to CCR7
  • Suitable for: IHC-P
  • Reacts with: Mouse, Human
  • Isotype: IgG

こちらの製品もご検討ください

二次抗体
Product image
Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
一次抗体
Product image
Anti-CD4 antibody [EPR19514] (ab183685)
一次抗体
Product image
Anti-CD4 antibody (ab203034)

関連製品

製品の概要

  • 製品名

    Anti-CCR7 antibody
    CCR7 一次抗体 製品一覧
  • 製品の詳細

    Goat polyclonal to CCR7
  • 由来種

    Goat
  • アプリケーション

    適用あり: IHC-Pmore details
  • 種交差性

    交差種: Mouse, Human
  • 免疫原

    Synthetic peptide corresponding to Mouse CCR7 aa 2-22.
    Sequence:

    DPGKPRKNVLVVALLVIFQVC

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • ポジティブ・コントロール

    • IHC-P: Mouse and human spleen tissues.
  • 特記事項

    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.

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • バッファー

    Preservative: 0.1% Sodium azide
    Constituents: 0.1% BSA, 0.81% Sodium chloride, 0.21% Potassium phosphate
  • Concentration information loading...
  • 精製度

    Affinity purified
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • Immunology
    • Innate Immunity
    • Chemokines
    • Beta Chemokine Rec. (CCR)
    • Signal Transduction
    • Signaling Pathway
    • G Protein Signaling
    • GPCR

関連製品

  • Compatible Secondaries

    • Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
    • Donkey Anti-Goat IgG H&L (HRP) (ab205723)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Goat IgG, polyclonal - Isotype Control (ab37373)

アプリケーション

Our Abpromise guarantee covers the use of ab1657 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
IHC-P 1/250.

ターゲット情報

  • 機能

    Receptor for the MIP-3-beta chemokine. Probable mediator of EBV effects on B-lymphocytes or of normal lymphocyte functions.
  • 組織特異性

    Expressed in various lymphoid tissues and activated B- and T-lymphocytes, strongly up-regulated in B-cells infected with Epstein-Barr virus and T-cells infected with herpesvirus 6 or 7.
  • 配列類似性

    Belongs to the G-protein coupled receptor 1 family.
  • 細胞内局在

    Cell membrane.
  • Target information above from: UniProt accession P32248 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 参照データベース

    • Entrez Gene: 1236 Human
    • Entrez Gene: 12775 Mouse
    • Omim: 600242 Human
    • SwissProt: P32248 Human
    • SwissProt: P47774 Mouse
    • Unigene: 370036 Human
    • Unigene: 2932 Mouse
    • 別名

      • BLR 2 antibody
      • BLR2 antibody
      • C C chemokine receptor type 7 antibody
      • C C CKR 7 antibody
      • C-C chemokine receptor type 7 antibody
      • C-C CKR-7 antibody
      • CC chemokine receptor 7 antibody
      • CC chemokine receptor type 7 antibody
      • CC CKR 7 antibody
      • CC-CKR-7 antibody
      • CCCKR7 antibody
      • CCR 7 antibody
      • CCR-7 antibody
      • Ccr7 antibody
      • CCR7_HUMAN antibody
      • CD 197 antibody
      • CD197 antibody
      • CD197 antigen antibody
      • CDw197 antibody
      • Chemokine (C C motif) receptor 7 antibody
      • Chemokine (C C) receptor 7 antibody
      • Chemokine C C motif receptor 7 antibody
      • Chemokine C C receptor 7 antibody
      • Chemokine receptor 7 antibody
      • Chemokine receptor 7-like protein antibody
      • CMKBR7 antibody
      • EBI 1 antibody
      • EBI1 antibody
      • Ebi1h antibody
      • EBV Induced G Protein Coupled Receptor 1 antibody
      • EBV-induced G-protein coupled receptor 1 antibody
      • Epstein Barr virus induced G protein coupled receptor antibody
      • Epstein Barr virus induced gene 1 antibody
      • Epstein-Barr virus-induced G-protein coupled receptor 1 antibody
      • EVI 1 antibody
      • EVI1 antibody
      • Lymphocyte Specific G Protein Coupled Peptide Receptor antibody
      • MGC108519 antibody
      • MIP 3 beta receptor antibody
      • MIP-3 beta receptor antibody
      • MIP3 Beta Receptor antibody
      see all

    画像

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR7 antibody (ab1657)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR7 antibody (ab1657)
      ab1657, at a dilution of 1/250, staining CCR7 in mouse spleen section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR7 antibody (ab1657)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR7 antibody (ab1657)

      ab1657, at a dilution of 1/250, staining CCR7 in mouse spleen section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR7 antibody (ab1657)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR7 antibody (ab1657)

      ab1657, at a dilution of 1/250, staining CCR7 in human spleen section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

    プロトコール

    • Immunohistochemistry - Paraffin Embedded Sections
    • Published IHC protocol for ab1657

    Click here to view the general protocols

    データシートおよび資料

    • Datasheet
    • SDS
  • 参考文献 (9)

    ab1657 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab1657 は 9 報の論文で使用されています。

    • Chen YS  et al. Ursodeoxycholic Acid Regulates Hepatic Energy Homeostasis and White Adipose Tissue Macrophages Polarization in Leptin-Deficiency Obese Mice. Cells 8:N/A (2019). PubMed: 30884843
    • Yang J  et al. Enhanced activity of macrophage M1/M2 phenotypes in periodontitis. Arch Oral Biol 96:234-242 (2018). PubMed: 28351517
    • Salim SY  et al. CD83+CCR7- dendritic cells accumulate in the subepithelial dome and internalize translocated Escherichia coli HB101 in the Peyer's patches of ileal Crohn's disease. Am J Pathol 174:82-90 (2009). IHC-P ; Human . PubMed: 19095953
    • Pitkin L  et al. Expression of CC chemokine receptor 7 in tonsillar cancer predicts cervical nodal metastasis, systemic relapse and survival. Br J Cancer 97:670-7 (2007). IHC-P ; Human . PubMed: 17687340
    • Liu JX  et al. CCR7, CCR8, CCR9 and CCR10 in the mouse hippocampal CA1 area and the dentate gyrus during and after pilocarpine-induced status epilepticus. J Neurochem 100:1072-88 (2007). PubMed: 17181556
    • Cinalli RM  et al. T cell homeostasis requires G protein-coupled receptor-mediated access to trophic signals that promote growth and inhibit chemotaxis. Eur J Immunol 35:786-95 (2005). PubMed: 15719365
    • Walker SR  et al. Murine neuroblastoma attenuates dendritic cell cysteine cysteine receptor 7 (CCR7) expression. J Pediatr Surg 40:983-7 (2005). PubMed: 15991182
    • Gattinoni L  et al. Acquisition of full effector function in vitro paradoxically impairs the in vivo antitumor efficacy of adoptively transferred CD8+ T cells. J Clin Invest 115:1616-26 (2005). PubMed: 15931392
    • Tanaka K  et al. Intratumoral injection of immature dendritic cells enhances antitumor effect of hyperthermia using magnetic nanoparticles. Int J Cancer 116:624-33 (2005). PubMed: 15825167

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-10 of 15 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CCR7 antibody

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA, pH9.0
    Sample
    Mouse Tissue sections (spleen)
    Specification
    spleen
    Permeabilization
    No
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Nov 19 2014

    Immunohistochemistry (Frozen sections) abreview for Anti-CCR7 antibody

    Average
    Abreviews
    Abreviews
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (spleen, lymph node)
    Specification
    spleen, lymph node
    Fixative
    Acetone
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Mr. Jong Hyung Lim

    Verified customer

    投稿 Nov 12 2009

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CCR7 antibody

    Below Average
    Abreviews
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (tonsil)
    Specification
    tonsil
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated
    Blocking step
    methanol/hydrogen peroxide (and I also tried 10% BSA) as blocking agent for 5 minute(s) · Concentration: 2.5%
    Read More

    Abcam user community

    Verified customer

    投稿 May 30 2007

    Question

    Huw called regarding the suggestions made.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 12 2006

    Answer

    Thank you for your patience. I have been in touch with the source of this antibody once again and I have some additional information. Unfortunately at this time I cannot source an alternative lot. However, the originator mentioned the following valuable information. They mentioned that blocking with 2% rabbit serum should be done prior to incubation with the primary antibody. There is no need for dilution of the antibody in 7.5% rabbit serum. Normal rabbit sera contains low levels of auto antibodies to various antigens in their blood. Normal rabbit serum at such high concentration (7.5%) contains a low level of cross-reacting antibodies that may inhibit binding of goat anti-mCCR7. IgG levels in rabbit serum could be 20-30mg/mL. At 7.5% NRS, the IgG level would be 1.5mg/mL in solution combined with approximately 4 ug/mL of goat anti-mCCR7 antibody. I hope this information helps, if this does not improve the results that you are obtaining I would be happy to provide you with a credit note against this purchase.

    Read More

    Abcam Scientific Support

    Answered on Dec 19 2006

    Question

    BATCH NUMBER 231100 ORDER NUMBER L076642 DESCRIPTION OF THE PROBLEM no staining on control sections (FFPE section of human tonsil and spleen) SAMPLE formalin fixed, paraffin embedded human tonsil tissue PRIMARY ANTIBODY abcam ccr7 (ab1657) made 1/250 in 7.5% normal rabbit serum DETECTION METHOD Vector ABC elite, 30 min at room temp. prepared as per protocol POSITIVE AND NEGATIVE CONTROLS USED human tonsil sections ANTIBODY STORAGE CONDITIONS stored at -80C on arrival to department FIXATION OF SAMPLE 24-48 hours in neutral buffered formalin ANTIGEN RETRIEVAL 2 separate protocols were performed- neither approach showed any staining 1)pH6.0 citrate buffer heating in microwave 2)Commercially available target antigen retieval solution in a 98C water bath PERMEABILIZATION STEP none BLOCKING CONDITIONS 7.5% Normal rabbit serum made in PBS at room temperature for 30 mins diluent SECONDARY ANTIBODY biotin conjugated rabbit anti-goat 1:400, in PBS, at room, 30 mins incubation HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? 2 separte retieval method have been employed with no postive staining seen in tissue ADDITIONAL NOTES the detection system and retrieval protocols are currently used successfully for other diagnostic immunohistochemical markers

    Read More

    Abcam community

    Verified customer

    Asked on Dec 05 2006

    Answer

    Thank you for taking the time to complete our questionaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody (ab16527). I have checked the procedure used to test this antibody in the laboratory (I have copied this below). Your method follows this very closely, although there are a couple of suggestions I can make to optimise your results. 1. To ensure the sample is not overblocked, I can suggest using just 2% serum for blocking. 2. Can you confirm the constituents of the commercial antigen retreival solution you have used? This antibody has given good results with EDTA buffer heat mediated antigen retrieval for 20 minutes (followed by cooling for 20 minutes). I suggest trying this if you have not done so already. 3. You can increase the signal by increasing the antibody concentration. I suggest trying 1:100. TESTING PROCEDURE: 1.Block the endogenous peroxide by incubating the sections with 3% H2O2 for 15 minutes at room temperature (RT). 2.Wash the sections with PBS and incubate with 2% normal rabbit serum at room temperature to block the non-specific binding sites. 3. Wash the section with PBS and incubate with primary antibody at recommended dilution for 60 minutes at RT. 4. Wash the slides and incubate at RT with biotinylated rabbit anti-goat IgG H+L PUBLISHED PROCEDURE: The following link cites this protocol in the following publication: http://clincancerres.aacrjournals.org/cgi/content/full/11/5/1743 Immunohistochemical Staining. The avidin-biotin complex method was used to detect anti-CCR7, dilution 1:250 Formalin fixed and paraffin-embedded tissues were deparaffinized and subsequently microwaved in EDTA buffer. After preincubation with hydrogen peroxide, avidin/biotin blocking kit, and rabbit serum, the primary antibody was applied for 1 hour at room temperature. After incubation with the secondary antibody (rabbit anti-goat biotinylated), the avidin-biotin complex was added and the enzyme activity visualized with diaminobenzidine. Counterstaining was done with hematoxylin. I hope this information is helpful. Should the suggestions not improve the results, please do not hesitate to contact me again.

    Read More

    Abcam Scientific Support

    Answered on Dec 06 2006

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CCR7 antibody

    Poor
    Abreviews
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (spleen)
    Specification
    spleen
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated
    Blocking step
    Serum as blocking agent for 15 minute(s) · Concentration: 10%
    Read More

    Dr. Linda Svantesson

    Verified customer

    投稿 Nov 17 2006

    Question

    BATCH NUMBER 195970 ORDER NUMBER 163068 DESCRIPTION OF THE PROBLEM Non-specific staining, high background with no specific signal seen SAMPLE Human tonsil and mouse multi organ block (lung, liver, spleen) PRIMARY ANTIBODY Abcam goat anti-CCR7 1/250 or 1/500 in 1% BSA/PBS; incubated overnight at 4C; wash PBS 2 x 10 minutes DETECTION METHOD ABC Elite, Vector Labs - 30 minutes room temp, wash PBS 2 x 10 minutes; followed by DAB. POSITIVE AND NEGATIVE CONTROLS USED Human tonsil and mouse spleen ANTIBODY STORAGE CONDITIONS aliquoted and stored at -20C when received 7/26/06. Aliquots removed and stored at 4C while testing IHC protocols. FIXATION OF SAMPLE 10% NBF, paraffin embedded; have also tried acetone and 4% PF fixed, frozen mouse spleen ANTIGEN RETRIEVAL heat mediated with citrate buffer (paraffin sections only) in steamer or Biocare Decloaker PERMEABILIZATION STEP None BLOCKING CONDITIONS Avidin/biotin blocking (Vector kit 15 min ea step); 10% normal rabbit serum 30 minutes. SECONDARY ANTIBODY biotinylated anti-goat 1/300, Vector labs, 30 minutes at room temp; wash PBS 2 x 10 minutes HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? antibody concentration; tissue fixation

    Read More

    Abcam community

    Verified customer

    Asked on Aug 07 2006

    Answer

    Thank you for your enquiry. I am sorry to hear that you have been obtaining non-specific staining using this antibody. I have read your technical questionnaire and I have a few comments. Please can you detail the duration of antigen retrieval that you have been performing. We make the recommendation that antigen retrieval is performed for 20 minute as detailed on the antibody datasheet. My concern is that the tissue preparation is being compromised influencing the presentation of the epitope to the antibody thereby causing non-specific staining. I would also appreciate comments with regards whether you have tried no antibody control experiments to control for non-specific staining of this antibody. Secondary antibodies are often discounted as being causal of non-specific staining. I look forward to hearing from you.

    Read More

    Abcam Scientific Support

    Answered on Aug 09 2006

    Question

    Obtaining actual WB band size of 60KDa, whereas the predicted is 43KDa (SwissProt)

    Read More

    Abcam community

    Verified customer

    Asked on Feb 09 2006

    Answer

    Thank you for your enquiry. Further to our telephone conversation yesterday afternoon I have been in touch with the source of this antibody. They informed me that by western blotting an actual molecular weight of 43KDa was detected in the following publication. I will e-mail this reference in a follow up e-mail. Walker SR et al. Murine neuroblastoma attenuates dendritic cell cysteine cysteine receptor 7 (CCR7) expression. J Pediatr Surg 40:983-7 (2005). PubMed: 15991182 It may be useful to examine the western blotting approach that was employed in this publication. However, following close examination of this reference it is clear that they do not show a molecular weight standard and the band is cropped in the image, therefore making it difficult to know the ACTUAL migration of the band. Should you continue to have difficulty with this antibody I would encourage you to submit a technical questionnaire by clicking on the link below. This will better enable our technical team to determine the steps that you have taken to optimise this antibody. https://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=1657&mode=questionaire I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Feb 10 2006

    Question

    I have read your protocol and I don't know which is the difference between yours and mine. Please, Could you help me? Protocol: 1. Please describe the problem (high background, no staining etc). We have high background and no staining in limphocytes. 2. On what material are you testing the antibody in IHC? ? Species? ? Cell line? ? Tissue? Human skin. Paraffin embedded 3. How did you fix the samples? ? Ethanol, methanol ? Acetone ? Paraformaldehyde ? Other Formaldehide 4% (24-48 hours). 4. Did you apply antigen retrieval step? ? Enzymatic method ? Heat mediated technique ? Other -Microwave 2 times, 3 minutes at 400 W. Citrate pH 6. -Steamer 30 minutes EDTA, or TrisEDTA. -Pressures Cooker, citrate pH6, 1 minute. 5. How did you block the unspecific binding sites? BSA 3% 30 minutes 6. Primary antibody ? 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)? CCR4 Goat. 1:300, 1:100, 1:600. CCR7 Goat. 1:250, 1:500 7. Secondary antibody ? 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? Antigoat secondary antibody 1:400. 8. Background staining ? Please provide an image of your staining. 9. Which detection system did you use? SEcondary antibody antigoat (dako) and kwick antibody Shandon. 10. Did you apply positive and negative controls along with the samples? Please specify. None. 11. Optimization attempts ? How many times have you tried the IHC? ? Do you obtain the same results every time? ? What steps have you altered? 5 times. Always the same results.

    Read More

    Abcam community

    Verified customer

    Asked on Feb 24 2005

    Answer

    Thank you for your e-mail. We hope these answers are more helpful. 5. How did you block the unspecific binding sites? BSA 3% 30 minutes Instead, use 2% normal rabbit serum at room temperature to block the non-specific binding sites. 7. Secondary antibody ? 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? Antigoat secondary antibody 1:400. Try to apply an ABC signal enhancing step. Wash the slides and incubate at RT with biotinylated rabbit anti-goat IgG H+L at 1:200 dilution for 30 minutes. Wash the slides and incubate with HRP-Streptavidin 1:400 dilution for 30 minutes. Wash the slides and incubate with DAB for 5-7 minutes at RT. Wash slides with PBS and counter stain with hematoxylin.

    Read More

    Abcam Scientific Support

    Answered on Feb 25 2005

    Question

    1. Please describe the problem (high background, no staining etc). We have high background and no staining in limphocytes. 2. On what material are you testing the antibody in IHC? Human skin. Paraffin embedded 3. How did you fix the samples? ? Formaldehide 4% (24-48 hours). 4. Did you apply antigen retrieval step? -Microwave 2 times, 3 minutes at 400 W. Citrate pH 6. -Steamer 30 minutes EDTA, or TrisEDTA. -Pressures Cooker, citrate pH6, 1 minute. 5. How did you block the unspecific binding sites? BSA 3% 30 minutes 6. Primary antibody CCR7 Goat. 1:250, 1:500 9. Which detection system did you use? SEcondary antibody antigoat and kwick antibody. 10. Did you apply positive and negative controls along with the samples? Please specify. None. 11. Optimization attempts ? How many times have you tried the IHC? Do you obtain the same results every time? ? What steps have you altered? 5 times. Always the same results.

    Read More

    Abcam community

    Verified customer

    Asked on Feb 22 2005

    Answer

    We are very sorry you are having problems with your protocol. Here is the up to date detailed protocol: .

    Read More

    Abcam Scientific Support

    Answered on Feb 24 2005

    1-10 of 15 Abreviews or Q&A

    •  戻る
    • 1
    • 2
    • 次へ 

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

    技術情報や製品情報をメールで受け取る 登録
    製品・研究分野別
    • 癌
    • 循環器
    • 細胞生物学
    • 発生
    • エピジェネティクス&核内情報
    • 免疫学
    • メタボリズム
    • 感染症
    • ニューロサイエンス
    • シグナル伝達
    • ステムセル
    製品・タイプ別
    • 一次抗体
    • 二次抗体
    • バイオケミカルズ
    • アイソタイプ・コントロール
    • マルチカラー・セレクター
    • キット
    • ローディング・コントロール
    • ライゼート
    • ペプチド
    • タンパク質
    • スライド
    • タグ抗体・細胞マーカー
    • その他試薬
    サービス&サポート
    • サポート
    • よくあるお問い合わせ
    • プロトコール&トラブルシュート
    • ご購入に関して
    • お問い合わせフォーム
    • トレーニング
    アブカムについて
    • 企業ウェブサイト
    • 投資家の皆様へ
    • 企業情報
    • 採用情報
    • アブカムについて
    イベント
    • 展示会
    • カンファレンス
    ブログ

    ブログを読む

    International websites
    • abcam.com
    • abcam.cn

    Join with us

    • LinkedIn
    • facebook
    • Twitter
    • YouTube
    • Terms of sale
    • Website terms of use
    • Cookie policy
    • Privacy policy
    • Legal
    © 1998-2021 Abcam plc. All rights reserved.