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Biotin Anti-RFP antibody (ab34771)

Reviews (5)Q&A (26)References (75)
Western blot - Biotin Anti-RFP antibody (ab34771)
  • Western blot - Biotin Anti-RFP antibody (ab34771)

Key features and details

  • Biotin Rabbit polyclonal to RFP
  • Suitable for: WB
  • Conjugation: Biotin
  • Isotype: IgG

Overview

  • Product name

    Biotin Anti-RFP antibody
    See all RFP primary antibodies

  • Description

    Biotin Rabbit polyclonal to RFP

  • Host species

    Rabbit

  • Conjugation

    Biotin

  • Specificity

    ab34771 recognises RFP. The antibody is predicted to react with all RFP variants from Discosoma (including DsRed, TdTomato, mOrange etc.) based on sequence homology and polyclonality of the antibody. However, such crossreactivity has not been QC tested.

  • Tested applications

    Suitable for: WBmore details

  • Immunogen

    Fusion protein corresponding to RFP. Red Fluorescent Protein (RFP) fusion protein corresponding to the full length amino acid sequence (234aa) derived from the mushroom polyp coral Discosoma.

  • General notes

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

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.

  • Storage buffer

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

  • Purity

    Immunogen affinity purified

  • Purification notes

    This product was prepared from monospecific antiserum by immunoaffinity chromatography using Red Fluorescent Protein (Discosoma) coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. No reaction was observed against Human, Mouse or Rat serum proteins. ELISA was used to confirm specificity at less than 0.1% of target signal.

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab34771 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
WB
1/2000 - 1/10000. Detects a band of approximately 27 kDa.
Notes
WB
1/2000 - 1/10000. Detects a band of approximately 27 kDa.

Target

  • Relevance

    Fluorescent proteins have become a useful and ubiquitous tool for making chimeric proteins, where they function as a fluorescent protein tag. Typically they tolerate N- and C-terminal fusion to a broad variety of proteins. They have been expressed in most known cell types and are used as a noninvasive fluorescent marker in living cells and organisms. They enable a wide range of applications where they have functioned as a cell lineage tracer, reporter of gene expression, or as a measure of protein-protein interactions.

  • Alternative names

    • DsRed antibody
    • GFP like chromoprotein antibody
    • red fluorescent protein antibody
    • RFP antibody

Images

  • Western blot - Biotin Anti-RFP antibody (ab34771)
    Western blot - Biotin Anti-RFP antibody (ab34771)
    All lanes : Biotin Anti-RFP antibody (ab34771) at 1/2500 dilution

    Lane 1 : 10 µg of HeLa cell
    extract containing GFP
    recombinant protein
    Lane 2 : 10 µg
    of a HeLa lysate containing RFP recombinant protein.


    Secondary
    All lanes : IRDye™800 conjugated Goat anti-Rabbit IgG at 1/5000 dilution

    Observed band size: 27 kDa why is the actual band size different from the predicted?



    IRDye™800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR.
  • Western blot - Biotin Anti-RFP antibody (ab34771)
    Western blot - Biotin Anti-RFP antibody (ab34771)
    Biotin Anti-RFP antibody (ab34771) at 1/1000 dilution
    Secondary
    Peroxidase streptavidin secondary antibody at 1/40000 dilution

References (75)

Publishing research using ab34771? Please let us know so that we can cite the reference in this datasheet.

ab34771 has been referenced in 75 publications.

  • Thorsen AS  et al. Heterogeneity in clone dynamics within and adjacent to intestinal tumours identified by Dre-mediated lineage tracing. Dis Model Mech 14:N/A (2021). PubMed: 33093165
  • Rhodes J  et al. Perception of a divergent family of phytocytokines by the Arabidopsis receptor kinase MIK2. Nat Commun 12:705 (2021). PubMed: 33514716
  • Brommer B  et al. Improving hindlimb locomotor function by Non-invasive AAV-mediated manipulations of propriospinal neurons in mice with complete spinal cord injury. Nat Commun 12:781 (2021). PubMed: 33536416
  • Grau N  et al. Spatiotemporally controlled induction of gene expression in vivo allows tracking the fate of tumor cells that traffic through the lymphatics. Int J Cancer 147:1190-1198 (2020). PubMed: 31675122
  • Kwon J  et al. Bright ligand-activatable fluorescent protein for high-quality multicolor live-cell super-resolution microscopy. Nat Commun 11:273 (2020). PubMed: 31937765
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 31 Abreviews or Q&A

Immunoprecipitation abreview for Anti-RFP antibody (Biotin)

 Good
Abreviews
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (293T)
Total protein in input
100000 cells
Immuno-precipitation step
Other - Dynabeads
Specification
293T
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 01 2019

Question

Inquiry: Dear Abcam supporters, I am looking for an antibody which could correspond to a dsred transgenic zebrafish, Tg(sox17:dsred) for confocal projection. I am wondering which one is more suitable for me, ab62341,ab124754,or ab34771. Looking forward to your reply.

Read More
Answer

Thank you for contacting us.


Only ab34771 is predicted toreact with DsRed based on sequence homology and polyclonality of the antibody.


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

Question

Dear abcam team,

We have recently ordered the Anti-RFP antibody (Biotin) (ab34771) in order to detect the RFP variant tdTomato expressed under an endogenous promoter. We have tried it on paraffin embedded sections. The datasheet said that:"IHC-P: 1/400. Perform heat mediated antigen retrieval before commencing with IHC" Could you provide me with some more detailed information regarding the heat mediated antigen retrieval? Could you also provide me with the buffer conditions?
Thanks

Read More
Answer

Thank you for contacting us.

The buffer we have used is Sodium citrate buffer 6.0 pH. The heat mediated antigen retrieval method should be optimized by the end user.

We recommend incubating the sections for 5,10, or 15 minutes in citrate buffer in 700 watt Microwave. In case of pressure cooker at high pressure 2 minutes incubation would be enough.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Question

is same antibody available but without biotin?

Read More
Answer


The unconjugated antibody is ab124754.
https://www.abcam.com/rfp-antibody-ab124754.html


Read More

Question

Dear xxxx,

Here are some answers for the questions that you asked:

1- Please find attach the image of the blot when using anti-RFP (ab34771) as Image_1.

2- The secondary antibody used is the HRP conjugated anti-rabbit IgG from xxxx. This secondary antibody is used by all the lab members and we never had a problem with its results.

3- The diluent that was used for the ab62341 is the 5% milk in TBS-Tween 0.1%, but when the blocking was done with BSA 3% (diluted in TBS-Tween 0.1%), this antibody was diluted in BSA. Membrane and antibody were incubated ON at 4°C.

4- Washing steps were done with TBS-T 0.1% for at least 15 min per wash, 3 times at RT. Note that the box where the membrane was kept to be washed was always covered.

5- Please find attach the image of the blot when using different blocking solutions, as Image_2.

6- All the images that were sent to you are with the lowest exposure time that we did.

I hope that these answers will help in your investigation. If you need more information, please let me know.

Best regards,

Read More
Answer

Thank you for your reply and the detailed information provided. This will be very useful to investigate this case.

It seems that regardless of the blocking conditions used, with the antibody ab62341 you are seeing very high background. This is quite blotchy, leading me to think that there are particles in one of the buffers which is sticking non-specifically to the membrane. It could be that the exact origin of this could be ascertained by testing all the different buffers, but as I suspect you have been using the same blocking and wash buffers for the ab34771 (where this background was not observed) and ab62341 I think further investigation would not be warranted.

It may be that this blotchyness could be removed by filtering your solutions prior to their use. However, I can understand that you have already put a lot of effort into optimising this antibody. I would therefore be happy to provide a replacement vial of (ab62341), or if you would prefer, a different antibody altogether. I would also be able to arrange a credit note or refund if this is preferable to you.

In order to arrange this, could you please provide me with the order number of ab62341 (or if you do not have this information, the delivery address)?

I look forward to receiving your reply.

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Question

thanks again for your email and your support. We will submit the
Abreview once we have the data ready for you. The only thing I would
like to draw your attention to is the expiration date of the discount
code in your message. It says 04/04/12. Am I right to assume that you
meant 04/04/13 instead?
Best regards,

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Answer

Thank you for your email.

You are right; I meant the 04/04/13 of course. I can reassure you that the code is valid until 04/04/13.

I wish you good luck with your experiment and look forward to hearing about your experience with this antibody in Flow Cytometry.

Read More

Question

thank you very much for your email. We would like to make use of your
offer regarding the AbTrial for the RFP (Biotin) antibody ab34771.
Just to understand this correctly, when we order the antibody now we
don't need to mention the code you will send us. Only when we order
something after submitting the Abreview we will need to provide it in
order to get the discount of 410 Euros. Is that correct?
Thank you very much for your help.
Best regards,

Read More
Answer

DISCOUNT CODE: ABTRIAL-xxxxx
Expiration date: 04/04/12
Value: €410.00

I am very pleased to hear you would like to accept our offer and test ab34771 in Flow Cytometry. This code will give you €410.00 off your next order before the expiration date. To redeem this offer, please submit an Abreview for Flow cytometry 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.


It is correct, that you do not have to mention the code with your order of ab34771. The code will be activated by submitting the Abreview and needs to be mentioned with the next order.

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/abtrial.

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Question

I am a colleague of Nicole and we would both work with the RFP
antibody, in case we are going to order one.
Firstly, we would like to know, if the AbTrail offer also applies on
the biotinylated Anti-RFP antibody (ab34771)? We would use it in flow
cytometry with some secondary streptavidin antibody. Secondly, what
would be the value of the discount for the next order, if we go for
the AbTrial option?
Thanks a lot for your help!
Best regards,

Read More
Answer

Thank you for your answer.

I would be pleased to offer you the AbTrial for the ab34771 to use this antibody in FlowCytometry. If you decide to order this antibody please let me now in advance so that I can provide you with the code.

You will receive the whole value of the antibody you are testing to order a new product. That means, if you decide to test the ab34771 you will receive a code with a value of €410.00. The code will be activated after you submit the Abreview.

Please let me know about your decision. I look forward hearing from you.

Read More

Question

Hi,
I'm looking for an anti-mRFP antibody that is suitable to be used in
flow cytometry. I saw that you have one that is FITC conjugated (#
ab34764) but unfortunately I also have GFP expression in my cells and
can't use this one. Do you use the same antibody clone for the
biotinylated anti-RFP antibody (ab34771)? So maybe it would be
possible to use this one? Do you have any information whether it could
work in flow cytometry or would it even be possible to get a small
sample to test this? Of cause you could also offer other suitable
anti-mRFP antibodies.
Thanks and kind regards

Read More
Answer

Thank you for contacting us.

In this case I would recommend to use the Anti-RFP antibody (ab62341). If it is suitable for your experiment you could use a secondary antibody to detect the Anti RFP antibody.

If the conjugate needs to be on the primary antibody I would recommend to have a look at our conjugation kits https://www.abcam.com/index.html?pageconfig=resource&rid=13148.

Because the Anti-RFP antibody (ab62341) has not been tested for flow cytometry I would am pleased to offer you the AbTrial for the use of this product in flow cytometry. By participating in our AbTrial program you can use our products in an untested application or species without financial risk.

Simply follow these easy steps below to apply for our AbTrial Program:

1. Reply to this email, letting me know you are interested in testing this product.

2. I will email you an inactive personal discount code for the value of the product.

3. Purchase and test the product at the regular price.

4. Submit your results, including your discount code in the additional notes section of your Abreview.

5. Once the Abreview is submitted, the discount code will become active.

6. Apply your discount code on your next order to receive that value off.

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/abtrial.

I hope I could be of help and I look forward hearing from you about your decision regarding the AbTrial offer.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

Read More

Question

Thanks very much for your reply. I just have some quick question. Is the antibody tested in transgenic DSR mice? And whether it will react to the GFP? I'm looking forward your reply. Thanks very much.

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Answer

Thank you for your reply.

We attempt to list all the information that we have about any of our products on the datasheet provided the information is not commercially senstive. Currently we are not aware of ab34771 being tested in transgenic DsR mice. Ab34771 is predicted to react with all RFP variants (including DsRed, TdTomato, mOrange etc.) based on sequence homology and polyclonality of the antibody. However, such crossreactivity has not been QC tested.

The product has been tested against GFP as shown in the Western blot image on the datasheet. In this experimentHeLa cell extract containing GFP recombinant protein andHeLa lysate containing RFP recombinant protein where compared. There was no reaction against GFP.

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

1-10 of 31 Abreviews or Q&A

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