山羊抗亚美尼亚仓鼠IgG H&L (Biotin) (ab5744)

Submit a review Q&A (4)References (8)

Key features and details

  • Goat Anti-Armenian hamster IgG H&L (Biotin)
  • Conjugation: Biotin
  • Host species: Goat
  • Isotype: IgG
  • Suitable for: WB, ICC/IF, Dot blot, ELISA, Immunomicroscopy, IHC-P

概述

  • 产品名称

    山羊抗亚美尼亚仓鼠IgG H&L (Biotin)
    参阅全部 IgG 二抗

  • 宿主

    Goat

  • 靶标种属

    Armenian hamster

  • 特异性

    Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-biotin, anti-Goat Serum, Armenian Hamster IgG and Armenian Hamster Serum. Cross reactivity against mouse has not been determined.

  • 经测试应用

    适用于: WB, ICC/IF, Dot blot, ELISA, Immunomicroscopy, IHC-Pmore details

  • 免疫原

    Armenian Hamster IgG whole molecule

  • 偶联物

    Biotin

性能

  • 形式

    Liquid

  • 存放说明

    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.

  • 存储溶液

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride, 1% BSA
  • Concentration information loading...

  • 纯化说明

    This product was prepared from monospecific antiserum by immunoaffinity chromatography using Armenian Hamster IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities.

  • 共轭说明

    Biotinamidocaproate N-Hydroxysuccinimide Ester (BAC) Biotin/Protein Ratio: 10-20 BAC molecules per Goat IgG molecule.

  • 克隆

    多克隆

  • 同种型

    IgG

  • 研究领域

应用

Our Abpromise guarantee covers the use of ab5744 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 1/2000 - 1/10000.
ICC/IF 1/1000 - 1/5000.
Dot blot Use at an assay dependent dilution.
ELISA 1/250000.
Immunomicroscopy Use at an assay dependent dilution.
IHC-P Use at an assay dependent dilution.

图片

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Armenian hamster IgG H&L (Biotin) (ab5744)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Armenian hamster IgG H&L (Biotin) (ab5744)
    Ab5744 was used at dilution 1/500 with the primary antibody ab33483 in IHC-P. See the review on ab33483.

实验方案

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

文献 (8)

发表研究结果有使用 ab5744？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab5744 被引用在 8 文献中.

  • Zheng J  et al. Evaluation of metastatic niches in distant organs after surgical removal of tumor-bearing lymph nodes. BMC Cancer 18:608 (2018). PubMed: 29848296
  • Dinh CH  et al. Bardoxolone Methyl Prevents High-Fat Diet-Induced Colon Inflammation in Mice. J Histochem Cytochem 64:237-55 (2016). PubMed: 26920068
  • Onodera K  et al. GATA2 regulates dendritic cell differentiation. Blood 128:508-18 (2016). PubMed: 27259979
  • Dinh CH  et al. Bardoxolone Methyl Prevents Mesenteric Fat Deposition and Inflammation in High-Fat Diet Mice. ScientificWorldJournal 2015:549352 (2015). IHC-P . PubMed: 26618193
  • Dinh CH  et al. Bardoxolone Methyl Prevents Fat Deposition and Inflammation in Brown Adipose Tissue and Enhances Sympathetic Activity in Mice Fed a High-Fat Diet. Nutrients 7:4705-23 (2015). IHC-P . PubMed: 26066016
  • Accart N  et al. Revisiting fixation and embedding techniques for optimal detection of dendritic cell subsets in tissues. J Histochem Cytochem 62:661-71 (2014). PubMed: 24874853
  • Yu P  et al. Diet-induced obesity drives negative mouse vein graft wall remodeling. J Vasc Surg N/A:N/A (2013). PubMed: 23876511
  • Dahllöf MS  et al. The lysine deacetylase inhibitor Givinostat inhibits ß-cell IL-1ß induced IL-1ß transcription and processing. Islets 4:417-22 (0). PubMed: 23486342

客户评价及客户问答

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1-4 of 4 Abreviews or Q&A

Question

What is the appropriate secondary antibody for use with this prodict in IHC-P?

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Answer

The appropriate secondary antibody for use with this product will be a Secondary Antibody to Armenian Hamster IgG such as ab5744.

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Question

I apologize it was so long for me to get back to you.  We have decided to go ahead with the antibodies you suggested, ab33483, ab6640 and ab8918.  I believe it will be difficult to use all three on a single tissue sample so we are going to proceed with serial sectioning immunohistochemistry instead of fluorescent staining, hopefully with the paraffin embedded sections although I saw that the mannose receptor suggested against this.  I believe biotin secondaries are the most common for these applications?  Again I am pretty new to this procedure.  So we will need to order one for hamster, rat, and mouse if you may recommend them.  Also, if you have any opinions on a better conjugation choice or procedure choice feel free to suggest.  Thanks for your help! any way.

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Answer

Thank you for contacting us. I am very glad to hear that you will be using our products in your experiments. I can gladly recommend three biotin labeled antibodies to suit your needs and have placed links for biotin labeled antibodies below. I would like to offer a second option which will allow you to perform these experiments but will allow you future flexibility as well. I recommend purchase of unlabeled secondary antibodies and labeling these with the marker of your choice (biotin, fluorochrome) using our EasyLink direct conjugation kits. In this way you will be able to perform serial stains using a single primary/secondary combination and will be able to optimize and trust them in this assay. You will also have the ability to use these same antibodies, apply different labels and use them in different assays such as a double or triple staining with fluorescent tags, western blot with HRP or other assays. Our EasyLink antibody conjugation kits provide asimple and quick linking process whichis part of a new generation of conjugation procedures. These eliminate many of the difficulties which were common in previous generation of conjugation kits andallow a quick and easy process to conjugate an antibody of your choice, in whatever function you need. It has the advantage of being aone-stepantibody labeling method withno separation steps. Further information about our EasyLink kits may be found at the following links: Landing Page:https://www.abcam.com/index.html?pageconfig=resource&rid=13148&source=pagetrap&viapagetrap=easylink FAQs:https://www.abcam.com/index.html?pageconfig=resource&rid=13156 The unconjugated antibodies which I would recommend would be: ab98800, anti-mouse, unconjugated:https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG-H-L-pre-adsorbed-ab98800.html ab98383, anti-rat, unconjugated:https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Rat-IgG-H-L-pre-adsorbed-ab98383.html ab5738, anti-Armenian hamster, unconjugated:https://www.abcam.com/Armenian-Hamster-IgG-secondary-antibody-H-L-ab5738.html For your initial biotin labeling I would recommend one of the following EasyLink Biotin (type A) labeling kits: 3 x10ug size:https://www.abcam.com/EasyLink-Biotin-type-A-Conjugation-Kit-3-x-10%C2%B5g-Biotin-ab102865.html 3 x 100ug size:https://www.abcam.com/EasyLink-Biotin-type-A-Conjugation-Kit-3-x-100%C2%B5g-Biotin-ab102864.html The pre-conjugated biotin labeled antibodies can be found at the following links: ab98802 anti-mouse, biotin:https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG-H-L-Biotin-pre-adsorbed-ab98802.html ab98414 anti-rat, biotin:https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Rat-IgG-H-L-Biotin-pre-adsorbed-ab98414.html ab5744 anti-Armenian hamster, biotin:https://www.abcam.com/Armenian-Hamster-IgG-secondary-antibody-H-L-ab5744.html Finally, as the anti-mannose receptor antibody has not yet been tested in IHC-P, I would like to offer to you enrollment in our 100% Abreview testing discount program. DISCOUNT CODE: XXXXXXX Expiration date: XXXXX There is no obligation for you to participate in this program; however should you wish to participate this code will give you: 1 free PRIMARY ANTIBODY OR VALUE OFF OF ORDER before the expiration date. To redeem this offer, please submit an Abreview for IHC-P with ab8918 and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. Use our products? Submit an Abreview. Earn rewards! https://www.abcam.com/abreviews

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Question

We generally use the Vector Laboratories ABC kits for IHC. General protocol is as follows (minus wash steps) 1. Frozen sections (mouse spleen) are fixed in cold acetone for 15 min. 2. Endogenous peroxidase blocked with H2O2 in methanol. 3. Blocking stem using VectastainABC kit (for this antibody, normal goat serum) for 30 min RT. 4. Primary staining done at 1/100-1/10,000 dilution using CD11c (N418, from Ab33483) in 0.1% BSA/PBST overnight at 4 degrees. 5. Secondary staining using goat anti-armenian hamster-biotinylated (ab5744) 30 min RT. Tried 1/200, 1/300 in PBST w/ diluted goat serum. Also tried 1/300 in 2% normal mouse serum (abcam). 6. Vectastain ABC reagent (peroxidase), 30 min RT. Without primary, my secondary stains what seems to be the marginal area around the germinal centers.

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Answer

Thank you for this information.

I have noted that this primary/secondary antibody combination has been used and reviewed by another customer. Their Abreview includes a detailed protocol which you may wish to review. This review is at:https://www.abcam.com/index.html?datasheet=33483&tab=abreviews&intabreviewid=14724

It may be in your case that much of the background is caused by secondary antibody binding to
endogenous mouse IgG in the tissue being stained, and to Fc receptors on B cells, plasma cells and macrophages. As such it may be valuable to use some of the staining procedures for mouse on mouse staining. Blocking endogenous IgG may be done by incubating the sections with an unconjugated Fab fragment likeGoat F (ab')2 polyclonal Secondary Antibody to Mouse IgG - (Fab')2, pre-adsorbed (ab98754),after the normal blocking step.More details may be found at this link:
https://www.abcam.com/index.html?pageconfig=resource&rid=11465

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

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Question

Inquiry: There is considerable cross-reactivity with the anti-hamster IgG antibody with mouse IgG. This is something that should be listed on your product page. Do you have an anti-hamster secondary that has less cross-reactivity with mice?

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Answer

Thank you for contacting us.

This product is not tested against or cross adsorbed against mouse. Since this is not a cross adsorbed secondary antibody, cross reactivity with other species may often be expected.

I have searched our catalog for other anti-Armenian Hamster IgG product which are cross absorbed by we currently do not offer any of these. I would very much like to help you to resolve this problem however. If you could please send me the details of your protocol I would be glad to review those and see if I might be able to make suggestions which will reduce background reactivity. Please include as much information as posibble in those notes.


I look forward to hearing from you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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