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  1. Link

    donkey-rabbit-igg-hl-dylight-488-preadsorbed-ab96919.pdf

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Immunology Immunoglobulins Heavy Chain IgG
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Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96919)

  • Datasheet
Reviews (1)Q&A (8)References (15)

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

Shipping info

Promotion Information

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。

Western blot - Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96919)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96919)
  • Immunohistochemistry (Frozen sections) - Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96919)
  • Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96919)

Key features and details

  • Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed
  • Conjugation: DyLight® 488. Ex: 493nm, Em: 518nm
  • Host species: Donkey
  • Isotype: IgG
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt

Conjugates logo Related conjugates and formulations

Alexa Fluor® 405 Alexa Fluor® 488 Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 680 Alexa Fluor® 750 Alexa Fluor® 790 Alkaline Phosphatase Biotin DyLight® 550 DyLight® 594 DyLight® 650 FITC Gold 10nm Gold 12nm Gold 18nm Gold 25nm Gold 6nm HRP IRDye® 680RD Texas Red ® TRITC Unconjugated Unconjugated

こちらの製品もご検討ください

標識
Product image
Alexa Fluor® 647 Conjugation Kit (Fast) - Lightning-Link® (ab269823)
ブロッキング
Product image
Normal Donkey Serum (ab7475)

関連製品

製品の概要

  • 製品名

    Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed
    IgG 二次抗体 製品一覧
  • 由来種

    Donkey
  • ターゲット生物種

    Rabbit
  • 特異性

    By immunoelectrophoresis and ELISA this antibody reacts specifically with Rabbit IgG and with light chains common to other Rabbit immunoglobulins. No antibody was detected against non immunoglobulin serum proteins. Reduced cross-reactivity to bovine, chicken, goat, horse, human, mouse, pig and rat was detected.

     

  • アプリケーション

    適用あり: WB, IHC-P, ICC/IF, Flow Cytmore details
  • 吸着処理血清


    Chicken, Cow, Goat, Horse, Human, Mouse, Pig, Rat
    To ensure minimal cross-reactivity, the antibody has been pre-adsorbed with serum proteins from the following species.
    more details
  • 標識

    DyLight® 488. Ex: 493nm, Em: 518nm

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Store at +4°C.
  • バッファー

    pH: 6.8
    Preservative: 0.09% Sodium azide
    Constituents: 0.2% BSA, PBS
  • Concentration information loading...
  • 精製度

    Immunogen affinity purified
  • 特記事項(精製)

    Antiserum was cross adsorbed using bovine, chicken, goat, human, mouse, pig and rat immunosorbents to remove cross reactive antibodies. This antibody was isolated by affinity chromatography using antigen coupled to agarose beads and conjugated to DyLight® 488.
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • Immunology
    • Immunoglobulins
    • Heavy Chain
    • IgG
    • Secondary antibodies
    • anti-Rabbit
    • IgG
    • Fluorophore
    • DyLight® 488

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab96919の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB
1/1000 - 1/20000. Predicted molecular weight: 36 kDa.
IHC-P
1/50 - 1/500.
ICC/IF (1)
1/50 - 1/500.
Flow Cyt
1/50 - 1/200.
特記事項
WB
1/1000 - 1/20000. Predicted molecular weight: 36 kDa.
IHC-P
1/50 - 1/500.
ICC/IF
1/50 - 1/500.
Flow Cyt
1/50 - 1/200.

画像

  • Western blot - Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96919)
    Western blot - Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96919)
    rabbit anti-actin antibody at 1 µg/ml + HeLa whole cell lysate at 30 µg

    Secondary
    Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96919) at 1/1000 dilution (Donkey anti-rabbit IgG heavy and light chain cross-adsorbed antibody)

    Predicted band size: 36 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96919)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96919)This image is courtesy of an abreview submitted by Carl Hobbs, Kings's College London, United Kingdom.
    Glial fibrillary acidic protein(GFAP) staining in mouse Formalin Fixed Paraffin Embedded sections. For antigen retrieval HIER in citrate buffer was performed. Donkey polyclonal Secondary Antibody to Rabbit IgG - H&L (DyLight® 488), pre-adsorbed (ab96919) was used as a secondary antibdoy at a dilution of 1/50.
  • Immunohistochemistry (Frozen sections) - Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96919)
    Immunohistochemistry (Frozen sections) - Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96919)

    E14 mouse brain sample was stained with primary antibody (ab66155). The secondary antibody used was ab96919 (1:200 dilution, incubate for 1 hour at room temperature). The blue background was from DAPI.

    This image is courtesy of an anonymous Abreview

  • Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96919)
    Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96919)
    Emission spectra of DyLight® fluorochromes available in our catalog.
    Line colors represent the approximate visible colors of the wavelength maxima.

プロトコール

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

データシートおよび資料

  • Datasheet download

    Download

参考文献 (15)

ab96919 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。

ab96919 は 15 報の論文で使用されています。

  • Matthews AT  et al. Doxorubicin-Induced Fetal Mesangial Cell Death Occurs Independently of TRPC6 Channel Upregulation but Involves Mitochondrial Generation of Reactive Oxygen Species. Int J Mol Sci 22:N/A (2021). PubMed: 34299212
  • Li Q  et al. M1 Macrophage-Derived Nanovesicles Repolarize M2 Macrophages for Inhibiting the Development of Endometriosis. Front Immunol 12:707784 (2021). PubMed: 34354711
  • Ogino R  et al. Adipose-Derived Stem Cells Promote Intussusceptive Lymphangiogenesis by Restricting Dermal Fibrosis in Irradiated Tissue of Mice. Int J Mol Sci 21:N/A (2020). PubMed: 32485955
  • Jara C  et al. Microencapsulation of cellular aggregates composed of differentiated insulin and glucagon-producing cells from human mesenchymal stem cells derived from adipose tissue. Diabetol Metab Syndr 12:66 (2020). PubMed: 32774470
  • Si J  et al. Antiepileptic effects of exogenous ß-hydroxybutyrate on kainic acid-induced epilepsy. Exp Ther Med 20:177 (2020). PubMed: 33101467
View all Publications for this product

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

1-9 of 9 Abreviews or Q&A

Donkey Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
IF was performed on mouse cell-line. Fixation with 4% PFA for 1hr at RT, followed by blocking with 3% FCS solution. Primary antibody used from other company, diluted 1:1000 in blocking solution. Incubated for 16hr, 4oC. Secondary antibody diluted 1:250 in blocking solution, incubated for 1hr at room temperature.

Abcam user community

Verified customer

投稿 Jul 29 2021

Question

Buenos días ,
Me gustaría que me mandases el código para beneficiarme de la promoción de Abreviews. Tenemos decidido adquirir el anticuerpo primario del cual hemos hablado (ab5794) así como del decundario que me has sugerido (ab96919). En cuanto a la promoción del anticuerbo rabbit monoclonal la idea es adquirir el ab39370 Anti-Netrin 1 antibody.
En cuanto me envíes el código hago el pedido formal vía e-mail a la direción que me indiques.
Muchas gracias,

Read More

Abcam community

Verified customer

Asked on Nov 19 2012

Answer

Tal y como hemos quedado, he creado un código de descuento: XXXXXX para testar ab5794 en células de gato mediante ICC.

El código caduca el día 20-Marzo-2013.

Este código tiene un valor de €370 si se realiza la compra antes de la fecha de caducidad. Para beneficiarse de esta oferta, por favor, mándanos los resultados mediante una Abreview obtenidos al testar el anticuerpo. Incluye por favor en los “Additional Comments” el código indicado, para que nosotros sepamos a qué corresponde dicha publicación.

Para más información acerca de publicar un Abreview, por favor, visita la página: https://www.abcam.com/Abreviews

Te recuerdo que publicamos resultados tanto positivos como negativos en las datasheet de los productos. El código se activará una vez que se haya publicado el Abreview, y puede canjearse mencionando el código de descuento proporcionado a la hora de hacer el pedido, bien sea por teléfono, por fax, o por email. ES IMPRESCINDIBLE INDICAR DICHO CODIGO PARA PODER BENEFICIARSE DEL DESCUENTO.

Las condiciones y términos de esta oferta las encontrarás en:

https://www.abcam.com/abtrial

En el pedido que hagas ahora, indica el código del anticuerpo primario (ab5794) el secundario que deseas adquirir (ab96919), y el del anticuerpo gratuito Rabbit monoclonal.

Me temo que el que me indicas en el mail no forma parte de la promoción, sin embargo, https://www.abcam.com/netrin-1-antibody-epr5428-ab126729.html también reconoce Netrin 1 y sí forma parte de la promoción. Si te inclinas finalmente por éste, indica su código en el pedido así como el código de la promoción para que se descuente su importe (RABMAB-XBSMG).

Si quieres que te envíe yo directamente el presupuesto con los tres productos, aplicando el descuento, indícamelo y estaré encantada de hacerlo.

Si tienes cualquier duda, por favor no dudes en contactarnos.

Read More

Abcam Scientific Support

Answered on Nov 19 2012

Question

One of your costumers intends to buy the following secondary antibodies for ICC/ IF, double staining:

1-) Donkey F(ab')2 polyclonal Secondary Antibody to Mouse IgG - H&L (DyLight® 594), preadsorbed (ab98768) - Orange/Red

2-) Donkey anti-Rabbit IgG H&L (DyLight® 488) secondary antibody (ab96919) - Green

But she is in doubt about which are the best filters to use for this analysis.

She has a Zeiss Axiophot microscope plus filters:

I 450-490 FT510 LP520
III BP546 FT580 LP590

See attached images.

Do you think she can use this microscope plus one of these filters for visualize DyLight 488 and Dylight 594 signals at once (double staining image)?

If she can't, which filter do you suggest for this kind of analysis?

Read More

Abcam community

Verified customer

Asked on Sep 27 2012

Answer

Thank you for contacting us.

I think this information would need to be obtained from the manufacturer of the microscope and filters that the customer is planning touse. They would be able to exactly tell the customer which wavelengths can be detected with these 2 filters.

The filter I 450-490 FT510 LP520 would probably work with the for 488 secondary antibody (ab96919), but I am not certain about the second one. The manufacturer would also be able to tell what other filters to use to make this work for the 594 secondary antibody (ab98768).

The numbers 488 and 594 refer to the excitation wavelength of the fluorophore. The emission wavelength are 519 nm (for the 488) and 617 nm (for the 594 fluorophore).

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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Abcam Scientific Support

Answered on Sep 27 2012

Question

use of ab96919 on tissue sections

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Abcam community

Verified customer

Asked on Jun 29 2012

Answer

Thank you for your call.

As discussed on the phone, the ab96919 can be used in a variety of protocols and applications. Please find here also the links to our online immunohistochemistry (IHC)protocols.

IHC protocol summary: https://www.abcam.com/index.html?pageconfig=resource&rid=11757

IHC frozen sections: https://www.abcam.com/index.html?pageconfig=resource&rid=11383

IHC paraffin staining protocol (detailed): https://www.abcam.com/index.html?pageconfig=resource&rid=11384

I hope this information is helpful. Please do not hesitate to call back, if you would like to discuss the protocols or the procedures in more detail.

I wish youa great day!

Read More

Abcam Scientific Support

Answered on Jun 29 2012

Question

Product code: 96919
Inquiry: The datasheet of ab96919 shows the image of Immunohistochemistry (Frozen sections) of E14 mouse brain but for some reason IHC-Fr does not appear as a tested application. Kindly advise if the customer will get Abpromise for IHC-Fr in this case? Thanks in advance for the clarification. Best Regards,

Read More

Abcam community

Verified customer

Asked on May 31 2012

Answer

DISCOUNT CODE: 42087
Expiration date: 28/05/2012
I am very pleased to hear you would like to accept our offer and test ab96919 in IHC-Fr. 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-Fr 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.

Read More

Abcam Scientific Support

Answered on May 31 2012

Question

Inquiry: The datasheet of ab96919 shows the image of Immunohistochemistry (Frozen sections) of E14 mouse brain but for some reason IHC-Fr does not appear as a tested application. Kindly advise if the customer will get Abpromise for IHC-Fr in this case? Thanks in advance for the clarification. Best Regards,

Read More

Abcam community

Verified customer

Asked on May 30 2012

Answer

Thank you for your enquiry and your interest in our products.
I can certainly offer a discount code for testing this secondary antibody for IHC on frozen sections.
Could you please pass your customer's contact details for our internal record and I can process the code for you as soon as possible.
I look forward to hearing from you soon.

Read More

Abcam Scientific Support

Answered on May 30 2012

Question

Dear All,



Sending you attached a list of the tendered items. You can see the products at page 3-5. Please give all of the items a check and let us know which items are best available with Abcam.



Early response requested.



Sincere regards,

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Abcam community

Verified customer

Asked on Apr 17 2012

Answer

Thank you for your inquiry.

We have a number of products that are comparable with the products on the list you sent us. For future inquiriesI would like to direct you to the search facility on our website (www.abcam.com).

Please enter the name of the protein into the search box at the top of the home page. A list of products will be generated which can be further refined using the filters on the left hand side of the page. Using these filters will help you to identify products that have been tested in the applications and species you are interested in.

Please find a list of the product code and the links to the respective datasheets below:

ab1012 (Anti-Histone H3 (tri methyl K4) antibody [mAbcam1012] - ChIP Grade)

https://www.abcam.com/Histone-H3-tri-methyl-K4-antibody-mAbcam1012-ChIP-Grade-ab1012.html (or use the following: https://www.abcam.com/Histone-H3-tri-methyl-K4-antibody-mAbcam1012-ChIP-Grade-ab1012.html).

ab12209 (Anti-Histone H3 (tri methyl K4) antibody [mAbcam12209] - ChIP Grade)

https://www.abcam.com/Histone-H3-tri-methyl-K4-antibody-mAbcam12209-ChIP-Grade-ab12209.html (or use the following: https://www.abcam.com/Histone-H3-tri-methyl-K4-antibody-mAbcam12209-ChIP-Grade-ab12209.html).

ab6002 (Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade)

https://www.abcam.com/Histone-H3-tri-methyl-K27-antibody-mAbcam-6002-ChIP-Grade-ab6002.html (or use the following: https://www.abcam.com/Histone-H3-tri-methyl-K27-antibody-mAbcam-6002-ChIP-Grade-ab6002.html).

ab124462 (Anti-Flag Tag antibody)

https://www.abcam.com/DDDDK-tag-antibody-Equivalent-to-FLAG-antibodies-from-Sigma-ab124462.html (or use the following: https://www.abcam.com/DDDDK-tag-antibody-Equivalent-to-FLAG-antibodies-from-Sigma-ab124462.html).

ab6939 (Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Cy3 ®), pre-adsorbed)

https://www.abcam.com/Goat-Rabbit-IgG-HL-Cy3--preadsorbed-ab6939.html (or use the following: https://www.abcam.com/Goat-Rabbit-IgG-HL-Cy3--preadsorbed-ab6939.html).

ab96899 (Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (DyLight® 488), pre-adsorbed)

https://www.abcam.com/Goat-Rabbit-IgG-HL-DyLight-488-preadsorbed-ab96899.html (or use the following: https://www.abcam.com/Goat-Rabbit-IgG-HL-DyLight-488-preadsorbed-ab96899.html).

ab96919 (Donkey anti-Rabbit IgG H&L (DyLight® 488) secondary antibody)

https://www.abcam.com/Donkey-Rabbit-IgG-HL-DyLight-488-preadsorbed-ab96919.html (or use the following: https://www.abcam.com/Donkey-Rabbit-IgG-HL-DyLight-488-preadsorbed-ab96919.html).

ab96879 (Goat polyclonal Secondary Antibody to Mouse IgG - H&L (DyLight® 488), pre-adsorbed)

https://www.abcam.com/Goat-Mouse-IgG-HL-DyLight-488-preadsorbed-ab96879.html (or use the following: https://www.abcam.com/Goat-Mouse-IgG-HL-DyLight-488-preadsorbed-ab96879.html).

ab96871 (Goat anti-Mouse IgG H&L (DyLight® 488) secondary antibody)

https://www.abcam.com/Goat-Mouse-IgG-HL-DyLight-488-ab96871.html (or use the following: https://www.abcam.com/Goat-Mouse-IgG-HL-DyLight-488-ab96871.html).

ab97265 (Goat anti-Mouse IgG Fc (HRP) secondary antibody)

https://www.abcam.com/Goat-Mouse-IgG-Fc-HRP-ab97265.html (or use the following: https://www.abcam.com/Goat-Mouse-IgG-Fc-HRP-ab97265.html).

ab7481 (Goat Serum)

https://www.abcam.com/Normal-Goat-Serum-ab7481.html (or use the following: https://www.abcam.com/Normal-Goat-Serum-ab7481.html).

ab7487 (Rabbit Serum (sterile))

https://www.abcam.com/Normal-Rabbit-Serum-Sterile-ab7487.html (or use the following: https://www.abcam.com/Normal-Rabbit-Serum-Sterile-ab7487.html).

ab46666 (Herring sperm ssDNA)

https://www.abcam.com/Herring-sperm-ssDNA-ab46666.html (or use the following: https://www.abcam.com/Herring-sperm-ssDNA-ab46666.html).

ab52579 (Ribonuclease A protein (Active))

https://www.abcam.com/Natural-cow-Ribonuclease-A-protein-ab52579.html (or use the following: https://www.abcam.com/Natural-cow-Ribonuclease-A-protein-ab52579.html).

ab73430 (DNase I protein (Active))

https://www.abcam.com/Recombinant-human-DNase-I-protein-Active-ab73430.html (or use the following: https://www.abcam.com/Recombinant-human-DNase-I-protein-Active-ab73430.html).

ab64220 (Proteinase K)

https://www.abcam.com/Proteinase-K-Antigen-Retrieval-Solution-ab64220.html (or use the following: https://www.abcam.com/Proteinase-K-Antigen-Retrieval-Solution-ab64220.html).

ab65621 (Protease Inhibitor Cocktail)

https://www.abcam.com/Protease-Inhibitor-Cocktail-ab65621.html (or use the following: https://www.abcam.com/Protease-Inhibitor-Cocktail-ab65621.html).

ab116028 (Prism Ultra Protein Ladder (10-245 kDa))

https://www.abcam.com/Prestained-Protein-Ladder-Broad-molecular-weight-10-245-kDa-ab116028.html (or use the following: https://www.abcam.com/Prestained-Protein-Ladder-Broad-molecular-weight-10-245-kDa-ab116028.html).


I hope this information is helpful to you. If you have specific questions about the products identified during the search, please do not hesitate to contact me.

Read More

Abcam Scientific Support

Answered on Apr 17 2012

Question

Dear Karin,

sorry, i thought i've already sent you the appendix, but there was something wrong with the address. Now here it is!

Best regards,

Read More

Abcam community

Verified customer

Asked on Jan 30 2012

Answer

Thank you for providing that extra information. Out of the secondaries used, the DyLight488 (ab96919) definitely seems to be producing more diffuse, non-nuclear staining compared to the other three. It may be that this could be reduced by optimising the blocking conditions and the antibody concentration. However, from the protocol you have used, I would expect the results to be better.

It may be that unfortunately you have received a faulty batch of this product. I am very sorry for the inconvenience this has caused.

If you would like, I can change the antibody for an alternative lot or I can offer an alternative antibody altogether (such as ab96895 or ab96899). These are both secondary antibodies to rabbit IgG with DyLight488 conjugation. If you would prefer a different conjugate I could accommodate this too. Alternatively I can provide a full refund.

In order to process any of these options I will need an order number on which the antibody was ordered (or an approximate date of the order and the delivery address).

I look forward to hearing how you would like to proceed.

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Abcam Scientific Support

Answered on Jan 30 2012

Question

LOT NUMBER GR63301-1 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM non-specific staining SAMPLE rat/brain PRIMARY ANTIBODY prim. AK: pCREB(rb) (Cell Signaling) 1:800 dilution: 5mg BSA in 990µl 0,05% Triton-X100 + 10µl NaN³ incubation: ˜ 36h at 4°C (3x5min PBS washing step) DETECTION METHOD fluorescence-microscopy POSITIVE AND NEGATIVE CONTROLS USED neg. control: withough prim. AK pos. control: prim. AK stained with Cy2 or Alexa488 ANTIBODY STORAGE CONDITIONS 4°C FIXATION OF SAMPLE 4% Paraformaldehyd, 0,1% Glutaraldehyd Perfusion for 1,15 h ANTIGEN RETRIEVAL - PERMEABILIZATION STEP (before 30min Glycinbuffer) 30min 0,1% Triton-X100 in PBS (3x5min PBS washing step) BLOCKING CONDITIONS 30min 1% Milkpowder-Dilution (3x5min PBS washing step) SECONDARY ANTIBODY sek. AK: a-rb DyLight488(dk) (Abcam) 1:200 dilution: 975µl 0,05% Triton-X100, 20µl Milkpowder-dilution, 5µl sek. AK incubation: 1h at ˜ 24°C (3x5min PBS washing step) 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? leaving out Milkpowder ADDITIONAL NOTES DyLight488 is fluorescent (checked it in it's clean form under the microscope), but does not bind the prim. AK with our protocol.

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Abcam community

Verified customer

Asked on Jan 27 2012

Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that the Donkey anti-Rabbit IgG H&L (DyLight® 488) secondary antibody (ab96919) 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 clarify a few points in order to understand the problems you have been having more fully.

1. You mention that the problem is non-specific staining, could you share the image obtained from the staining with me? Do you get the same staining with the "no-primary" control??

2. Do you have the catalogue number for the primary antibody used? Was this the same primary antibody used with the Cy2 and Alexa488 staining? Would you be able to share the images produced from these experiments?

These details will greatly help me to understand what may be going wrong.

In the event that a product 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. In order to process this I would need an order number or the date of the order and the delivery address in order to locate your order.

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

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Abcam Scientific Support

Answered on Jan 27 2012

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

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