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  1. Link

    goat-rabbit-igg-hl-dylight-488-preadsorbed-ab96899.pdf

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Immunology Immunoglobulins Heavy Chain IgG
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Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)

  • Datasheet
  • SDS
Submit a review Q&A (13)References (81)

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Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Western blot - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Flow Cytometry - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Flow Cytometry - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)

Key features and details

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

Conjugates logo Related conjugates and formulations

Agarose Alexa Fluor® 405 Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alexa Fluor® 680 Alexa Fluor® 680 Alexa Fluor® 750 Alexa Fluor® 790 Alkaline Phosphatase APC beta-galactosidase Biotin Cy2 ® Cy3 ® Cy3.5 ® Cy5 ® Cy5.5 ® DyLight® 550 DyLight® 594 DyLight® 650 FITC Glucose Oxidase Gold 12nm Gold 6nm HRP HRP polymer IRDye® 800CW PE Texas Red ® TRITC Unconjugated Unconjugated

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Overview

  • Product name

    Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed
    See all IgG secondary antibodies
  • Host species

    Goat
  • Target species

    Rabbit
  • Specificity

    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 IgG was detected.

     

  • Tested applications

    Suitable for: IHC-P, ICC/IF, Flow Cyt, WBmore details
  • Minimal
    cross-reactivity


    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
  • Conjugation

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C.
  • Storage buffer

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

    Immunogen affinity purified
  • Purification notes

    Antiserum was cross absorbed using bovine, chicken, horse, 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.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab96899 in the following tested applications.

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

Application Abreviews Notes
IHC-P
1/50 - 1/500.
ICC/IF
1/50 - 1/500.
Flow Cyt
1/500.
WB
1/1000 - 1/20000. Predicted molecular weight: 36 kDa.

5% non-fat dry milk in PBST or TBST is recommended for blocking and incubation of antibodies. BSA is not recommended.

Notes
IHC-P
1/50 - 1/500.
ICC/IF
1/50 - 1/500.
Flow Cyt
1/500.
WB
1/1000 - 1/20000. Predicted molecular weight: 36 kDa.

5% non-fat dry milk in PBST or TBST is recommended for blocking and incubation of antibodies. BSA is not recommended.

Images

  • Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
    Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)Han et al PLoS One. 2014 Jan 23;9(1):e86539. doi: 10.1371/journal.pone.0086539. eCollection 2014. Fig 5. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Effects of FZE on apoptotic ratio and apoptotic factors in RSC96 cells.

    Effects of FZE on translocation of CytoC and the levels of caspase9 and caspase3. The cells were fixed with 4% paraformaldehyde for 15 minutes at 20°C, permeated with 0.3% triton prior to being blocked in 1% BSA+2% normal goat serum for 30 min at 20°C. Samples were then incubated with primary antibody overnight at 4°C in PBS containing. ab96899 diluted at 1∶200 was used as the secondary antibody. Cell nucleus were counterstained with DAPI and showed blue. Mitochondria were labeled by Mito tracker and showed red.

  • Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
    Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)

    ICC/IF image of (ab3280) stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

    The cells were fixed in 100% methanol (5 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3280, 5 µg/ml) overnight at +4°C.

    The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) used at a 1/250 dilution for 1 h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.

  • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
    Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)

    Emission spectra of DyLight® fluorochromes available in our catalog.
    Line colors represent the approximate visible colors of the wavelength maxima.

  • Western blot - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
    Western blot - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
    All lanes : Cocktail of rabbit anti-Actin and mouse anti-hnRNP at 1 µg/ml

    All lanes :

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899) at 0.5 µg/ml (Cocktail of Dylight® 488-conjugated goat anti-rabbit ab96899 (blue) and Dylight® 680-conjugated goat anti-mouse (red))

    Predicted band size: 36 kDa


    Exposure time: 42 seconds
  • Flow Cytometry - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
    Flow Cytometry - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
    Overlay histogram showing A431 cells stained with unpurified ab124962 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab124962, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
    Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
    ab96379 staining MEK1 (phospho S298) in SK-N-SH cells treated with CNQX (ab120017), by ICC/IF. Decrease in MEK1 (phospho S298) expression correlates with increased concentration of CNQX, as described in literature.
    The cells were incubated at 37°C for 24h in media containing different concentrations of ab120017 (CNQX) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab96379 (1/100 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody.
  • Flow Cytometry - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
    Flow Cytometry - Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)

    Overlay histogram showing Raji cells stained with ab52922 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52922, 1/100) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Raji cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

Protocols

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

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (81)

Publishing research using ab96899? Please let us know so that we can cite the reference in this datasheet.

ab96899 has been referenced in 81 publications.

  • Zhang J  et al. Microrchidia family CW-type zinc finger 2 promotes the proliferation, invasion, migration and epithelial-mesenchymal transition of glioma by regulating PTEN/PI3K/AKT signaling via binding to N-myc downstream regulated gene 1 promoter. Int J Mol Med 49:N/A (2022). PubMed: 34913078
  • Wu N  et al. Essential role of MALAT1 in reducing traumatic brain injury. Neural Regen Res 17:1776-1784 (2022). PubMed: 35017438
  • Hu Y  et al. MiR-223 Promotes Tumor Progression via Targeting RhoB in Gastric Cancer. J Oncol 2022:6708871 (2022). PubMed: 35035482
  • Luo H  et al. Beclin-1 exerts protective effects against cerebral ischemia-reperfusion injury by promoting DNA damage repair through a non-autophagy-dependent regulatory mechanism. Int J Mol Med 49:N/A (2022). PubMed: 35266018
  • Zhou Y  et al. Effects and early diagnostic value of lncRNA H19 on sepsis-induced acute lung injury. Exp Ther Med 23:279 (2022). PubMed: 35317444
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-10 of 13 Abreviews or Q&A

Question

Bonjour,

Je travaille sur des cellules équines et je suis à la recherche d’un anticorps anti-Mucin5AC qui ne soit pas mouse.

Je viens de voir sur votre site que vous aviez un anticorps rabbit anti-mucin5AC (ab78660) mais malheureusement il n’est pas décrit comme croisant avec le cheval.



Serait-il possible d’obtenir une offre promotionnelle pour tester cet anticorps (ab78660) sur des cellules équines ainsi que pour le secondaire ab96899?



Très cordialement

Read More

Abcam community

Verified customer

Asked on Oct 10 2012

Answer

Merci de votre intérêt pour l'anti-Mucin5AC ab78660 et le secondaire ab96899.

En effet, l'anticorps ab78660 n’a pas encore été testé chez le cheval et nous n’offrons pas d’échantillon gratuit à des fins de tests préliminaires.
Je n'ai pas trouvé la séquence de la Mucin5AC du cheval et je n'ai donc pas pu déterminer l'homologie de séquence avec le peptide immunogène et les chances que cet anticorps fonctionne pour cette espèce. Disposez-vous de cette séquence?

Si l'homologie est bonne et si vous souhaitez tester ab78660 chez le cheval, il me sera possible de vous offrir une remise. Après soumission d’une Abreview avec vos résultats, un avoir de la valeur de l'anticorps testé vous sera offert.

Cette offre de test n'est valable que pour les anticorps primaires, pas pour les secondaires et donc pas pour le ab96899.

Etapes à suivre :

1. Nous confirmer que vous souhaitez tester ab78660 chez le cheval afin de recevoir le code de remise correspondant. Ce code doit être édité avant l’achat de ab78660

2. Acheter ab78660 par téléphone, fax ou internet (https://www.abcam.com)

3. Le tester chez le cheval

4. Nous faire part de vos résultats, positifs ou négatifs, grâce à notre système Abreview. Pour plus d’informations https://www.abcam.com/abreviews

5. Après soumission de votre Abreview, appeler notre service clientèle afin de passer votre prochaine commande grâce au code de réduction. Ce code de réduction est utilisable 120 jours après son édition et utilisable lors de l’achat d’un autre anticorps primaire.
Même si ab78660 ne pas fonctionne pas chez le cheval, après soumission de votre Abreview, vous recevrez quand même un avoir de la valeur de l'anticorps testé.
Termes et Conditions : https://www.abcam.com/collaborationdiscount.

N’hésitez pas à nous demander plus d’informations concernant cette offre promotionnelle. Je vous envoie dans un autre email un devis pour les 2 anticorps.

Read More

Abcam Scientific Support

Answered on Oct 10 2012

Question

Customer kindly contacted us to inquire about the immunogen sequence for ab56215.

Read More

Abcam community

Verified customer

Asked on Oct 02 2012

Answer

Thank you for contacting us and your interest in our products. The immunogen sequence used to raise theab56215 Anti-SLC4A4 antibodyis proprietary information. However, in order to aid your research I am able to share it with you. Please do not share or publish this information. It is strictly for your own use. The peptide sequence used is as follows: Abcam also carries secondary antibodies for use in Flow Cytometry and Direct Conjugation Kits. Our EasyLink kits providea simple and quick process to conjugate your primary antibodies with FITC. Ab102885 EasyLink FITC Conjugation Kit can be found at the following link: https://www.abcam.com/easylink-fitc-conjugation-kit-3-x-10%C2%B5g-fitc-ab102885.html OurGoat polyclonal Secondary Antibody to Rabbit IgG - H&L (DyLight® 488), pre-adsorbed can be found at: https://www.abcam.com/goat-polyclonal-secondary-antibody-to-rabbit-igg-h-l-dylight-488-pre-adsorbed-ab96899.html For a limited time we are offering our Cell Imaging Book with any secondary antibody order. Just include the codeCELL-TBVCQ when ordering. More information about this promotion and our Cell Imaging Book can be found at: https://www.abcam.com/index.html?pageconfig=resource&rid=14705 I hope this information has been of help to you. If you have any further questions, please do not hesitate to ask.

Read More

Abcam Scientific Support

Answered on Oct 02 2012

Question

Since neither ab28856 nor ab51059 is tested in IHC-Fr. We are thinking of testing ab28856 first.

Could you send me a discount code for testing ab28856 (Anti-NFkB p65 (phospho S536) antibody) only?

Read More

Abcam community

Verified customer

Asked on Jun 12 2012

Answer

DISCOUNT CODE: **********
Expiration date: *************

I am very pleased to hear you would like to accept our offer and test ab28856 inIHC-Fr. This code will give you: 1 free PRIMARY ANTIBODY 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 Jun 12 2012

Question

Testing discount info for ab28856 & ab51059 in IHC-Fr sections.

Read More

Abcam community

Verified customer

Asked on Jun 11 2012

Answer

Thank you very much for your interest in 28856 & ab51059.

To our knowledge, 28856 & ab51059 have not been tested in IHC-Fr. Therefore, I can offer a discount off a future purchase if you buy 28856 & ab51059 now, test it in IHC-Fr 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 28856 & ab51059 in IHC-Fr. I will then send a discount code. This code must be issued before purchasing 28856 & ab51059 so please wait for my reply before ordering.

2. Purchase 28856 & ab51059 either by phone, fax, or online (www.abcam.com).

3. Test it in IHC-Fr.

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 how to submit an Abreview, 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.
Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if 28856 & ab51059 turns out to be unsuitable for IHC-Fr 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.

Read More

Abcam Scientific Support

Answered on Jun 11 2012

Question

Thank you for your reply. I checked this product and it says that the target species is Rabbit while I want to apply it on Human tissue. so does it work on human tissue? if not can you please suggest another appropriate product.



Thanks

Read More

Abcam community

Verified customer

Asked on May 18 2012

Answer

Thank you for contacting us.

The ab292 is a rabbit antibody which target human collagen I. To detect this primary you would need secondary which is ab96899.

If your primary ab works in human then the secondary ab will work.

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

Read More

Abcam Scientific Support

Answered on May 18 2012

Question

Inquiry: I ordered an anti-collagen type1 from Abcam (Rabbit poly, ab292) and I want to apply Immunofluorescence experiment on HUMAN cornea tissue. I need to order now an anti mouse/rabbit? secondary antibody conjugated with AlexaFluor 488. So, first is it better to order it with anti mouse or anti rabbit, and can you help me in what product to choose because there are too many options. Thank you

Read More

Abcam community

Verified customer

Asked on May 02 2012

Answer

Thank you for contacting us.

Please buy ab96899;https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Rabbit-IgG-H-L-DyLight-488-pre-adsorbed-ab96899.html

The Dylight 488 conjugate is better than Alexa 488.

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

Read More

Abcam Scientific Support

Answered on May 02 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,

Read More

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.

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Abcam Scientific Support

Answered on Apr 17 2012

Question

Thank you very much! In this way I should be able to image the aggrecan in green and the collagen II in red, is this correct? Is there any kind of discount or special price for this package?

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Abcam community

Verified customer

Asked on Mar 13 2012

Answer

It is my pleasure to help.

In this particular group, aggrecan would fluoresce in green while collagen II would be red. We offer other dyes athttps://www.abcam.com/index.html?pageconfig=resource&rid=12956should you wish to change those colors.

We do offer discounts on purchases of five items or more and offer a number of promotions. Our customer service representatives will be able to give you more detailed information about our current promotions and discounts or you may follow the link here:https://www.abcam.com/index.html?pageconfig=resource&rid=14853

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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Abcam Scientific Support

Answered on Mar 13 2012

Question

Hello, I need to image collagen type II and aggrecan from mouse within the same samples. The laser that I have available are 458, 488, 514, 543, 633. Which combination of antibodies (primary and secondary) do you recommend that might work nicely together? Many thanks for your assistance.

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Abcam community

Verified customer

Asked on Mar 12 2012

Answer

Thank you for contacting us.

We presently have only 1 product against mouse aggrecan, ab36861. This is a rabbit polyclonal. In ICC and appropriate secondary would be anti-Rabbit IgG with a fluorescent tag which could be excited by your lasers. A product such as ab96899,Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (DyLight® 488),https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Rabbit-IgG-H-L-DyLight-488-pre-adsorbed-ab96899.html

We currently have 20 products against collagen II. However taking into account that this will have to be used in mouse tissue in conjunction with an anti-aggrecan rabbit polyclonal we can limit this to ab3092Anti-Collagen II antibody [5B2.5]. This is a mouse monoclonal antibody of isotype IgG2a that has been tested inIHC-Fr,IHC-P,WB and IF. An appropriate secondary should preferably be a F(ab) fragment as mouse on mouse staining can be difficult, please see this link for more information about mouse on mouse staining,https://www.abcam.com/index.html?pageconfig=resource&rid=11465. One such product would be ourGoat F(ab')2 polyclonal Secondary Antibody to Mouse IgG - (Fab')2 (DyLight® 550), pre-adsorbed,https://www.abcam.com/Goat-F-ab-2-polyclonal-Secondary-Antibody-to-Mouse-IgG-Fab-2-DyLight-550-pre-adsorbed-ab98758.html

Abcam also offers a number of ICC and IHC reagents. As both of the secondary antibodies listed above are raised in goat, we recommend using a 5-10% Goat serum as your blocking reagent and antibody incubation solution, ab7481https://www.abcam.com/Goat-Serum-ab7481.html. We, also offer buffers and antigen retrieval solutions,https://www.abcam.com/index.html?pageconfig=resource&rid=12854, as well as mounting media for fluorescent staining ab103748,https://www.abcam.com/BrightMount-Plus-25ml-Aqueous-Mounting-Medium-Anti-fading-for-Fluorescent-Staining-ab103748.html.


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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Abcam Scientific Support

Answered on Mar 12 2012

Question

I have run an IF-staining on bronchoalveolar cells (human: mainly alveolar macrophages and T lymphocytes; cytospin slides 60 000 cell per slide; sarcoidosis patients) with:

1- Anti-PPAR alpha antibody (ab61182) + Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (DyLight® 488), pre-adsorbed (ab96899) and you can check the results in the PDF attachment. I think it works. 4% formaldehyde for fixation, fish serum for blocking; saponin for permeabilisation.

My question: as a negative control I omitted the primary antibody and my question is that enough or do I have to include your immunizing Abs or another methodto be sure?

2- Simple (only antiCD4) and double staining (Anti PPAR and CD4),I have observed that Mouse monoclonal to CD4 (ab25804) + Rat monoclonal secondary Ab to Mouse IgG1 (PE) did not stainCD4 Tcells.

Also my friend tested these Ab in lung tissue and they did not work.

My Question: Why? Do recommend another solution? Or Can I get another Ab from you?

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Abcam community

Verified customer

Asked on Jan 20 2012

Answer

Thank you for contacting us.
To answer your questions:
1. Yes, a negative control can be ommiting the primary. Other options could be using an isotype control antibody that compares with the priamry, or the blocking peptide. There are various options, but it is up to each reseracher what they prefer to use.
PPAR alpha should stain the nucleus, and your PPAR alpha data do not seem to show that - from what I can see. The DAPI stain seems to dominate the image, and the green from Dylight 488 is difficult to see - which should be also in the nucleus. You might want to increase the amount of antibody being used (for the secondary, but maybe also the primary), or switch to a red color which would be more easy to see in contrast with the blue (if possible). Using less DAPI could help as well.
You also might want to use a stronger detergent than saponin (e.g. NP40 or TX-100) in order to permeabilize the nuclear membrane).
How long did you permeabilize for with 4%PFA?

2) As for the CD4 antibody, please let me what protocol you used for the double staining including antibody dilutions, sequence of staining, fixation step, blocking step etc.
I look forward to hear back from you.

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Abcam Scientific Support

Answered on Jan 21 2012

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