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  1. Link

    biotin-rfp-antibody-ab34771.pdf

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Tags & Cell Markers Fusion / Marker Proteins RFP
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Biotin Anti-RFP antibody (ab34771)

  • Datasheet
  • SDS
Reviews (5)Q&A (26)References (75)

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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

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Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795)
Protein
Product image
Recombinant RFP protein (ab51993)

View more associated products

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

    • Tags & Cell Markers
    • Fusion / Marker Proteins
    • RFP

Associated products

  • Isotype control

    • Biotin Rabbit IgG - Isotype Control (ab200208)
  • Recombinant Protein

    • Recombinant RFP protein (ab51993)
  • Related Products

    • Biotin Assay Kit (Colorimetric) (ab185441)

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

Protocols

  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

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

11-20 of 31 Abreviews or Q&A

Immunocytochemistry abreview for Anti-RFP antibody (Biotin)

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Mouse Cultured Cells (neuronal)
Specification
neuronal
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton-x 100
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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 Apr 17 2012

Question

Inquiry: Hello, I' m looking for anti-RFP, I'm wondering why you conjugate biotin on the primary antibody? If I want to do IHC & IF on mice mammary gland, which one is the best you recommendation? I'll be very appreciate your help, thanks a lot.

Read More

Abcam community

Verified customer

Asked on Sep 25 2012

Answer

Thank you very much for contacting us with your question.

Ab34771 is conjugated to biotin so that a biotin-streptavidinsystem can be used in the detection of the antibody, which will amplify the signal. To use this antibody in immunofluorescent staining in IHC, you would usestreptavidin conjugatedto a fluorophore instead of the secondary antibody step. We have several streptavidinconjugatesin our catalog that can be used with Ab34771, such as ab13622 which is conjugated to FITC-

https://www.abcam.com/Streptavidin-protein-FITC-ab136222.html

If you prefer an unconjugated RFP antibody to use with a secondary conjugated antibody, we also have Ab62341 which has been tested with frozen and paraffin-embedded tissue in IHC-

https://www.abcam.com/RFP-antibody-ab62341.html

Ab62341 has also been referenced in 10 Abreviews and 11 publications, so I think this will be a good choice for your staining.

I hope that this information will be useful, but please let me know if you have further questions and I'll be happy to help.

Read More

Abcam Scientific Support

Answered on Sep 25 2012

Question

Do you have the ab34771 immunogen sequence information?

Read More

Abcam community

Verified customer

Asked on Sep 18 2012

Answer

Thank you for your patience.



I have enquired with the product originator but unfortunately, the actual sequence information isproprietary and cannot be released. They did however mention that the sequence wasderived from thereference links below:


http://www.ncbi.nlm.nih.gov/protein/55976617?report=genbank&log$=prottop&blast_rank=2&RID=56D9X48A01N

http://www.uniprot.org/uniprot/Q9U6Y8

I hope this information will still be helpful to you.

All of our products are guaranteed to work in species and application as stated on the datasheet. Should you not get expected results, then please inform us within the guarantee period of 6 month (Abpromise). We will help troubleshoot the experiment and if the product is indeed faulty, we will be more than happy to offer a free replacement or refund.

I hope our Abpromise guarantee will give you the assurance you need to use this product in your research.

Read More

Abcam Scientific Support

Answered on Sep 18 2012

Question

I'm interested in your product (anti-RFP antibody (biotin)), and the product number is ab34771. Does it work with mCherry? What is the molecular weight of this antibody?

Read More

Abcam community

Verified customer

Asked on Aug 28 2012

Answer

Thank you for contacting Abcam regarding ab34771.


This anti-RFP antibody is designed to react with all RFP variants and thus will work with mCherry. Regarding the molecular weight of the antibody, it is160kDa plus the weight of the biotin (244 Da). The molecular weight of the band being detected will depend on the MW of the protein of interest, plus the weight of mCherry (˜29kDa).

I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

Read More

Abcam Scientific Support

Answered on Aug 28 2012

Question

Product code: 34771

Inquiry: Dear Abcam Representative, I am a researcher in a lab at Karolinska Institutet, in Sweden. I am interested in an antibody of yours for advanced single molecule trafficking, but would like to test for binding before I invest in a sizable order.
This would be the first time studies of RFP tagged proteins have been monitored in real-time in brain cells. If possible to get a small aliquot for testing, I would really like to try out your Anti-RFP antibody (Biotin) (ab34771).
It has been used successfully for various other labeling methods, so I have hope that it will also effectively label for advanced live tracking.
This would be a very attractive technique with many applications and for any membrane proteins. Please contact me by mail if there is any information that would be helpful in accessing a test sample. Thank you kindly for your assistance, -

Read More

Abcam community

Verified customer

Asked on Jul 11 2012

Answer

Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. I am unsure what application your experiment belongs to. Maybe Immunomicroscopy(IM)?When on the datasheet, pleasereview the link"show applications key" (right hand side) next to Applications. Which application is the best match?

I will be happy to include you in our testing discount program.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

Read More

Abcam Scientific Support

Answered on Jul 11 2012

Question

RFP detected in IHC-Fr, ICC/IF
did not detect dsRed1 and dsRed2 - no signal
lot GR44251-4

Was Ab tested on dsRed1 and 2?
Was Ab tested with avidin (to show that biotin is functional)? Any data in this regard?

Read More

Abcam community

Verified customer

Asked on May 23 2012

Answer

According to our records, ab34771 was proving difficult to use in IHC and ICC and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced.

For our records, could you please send me a few more details about the staining that did not work (e.g. Antibody dilution, blocking conditions, sample type, detection systems used)? That would be of great help. Thank you.

In order to resolve this issue, I'd be happy to offer you a replacement, credit or refund if the antibody was purchased within the last 6 months. Please let me know your order or PO number in that regard as well as which option would be best for you.

If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

I wish you the best of luck with your research and look forward to a reply.

Read More

Abcam Scientific Support

Answered on May 23 2012

Question

RFP detected in IHC-Fr, ICC/IF
did not detect dsRed1 and dsRed2 - no signal
lot GR44251-4

Was Ab tested on dsRed1 and 2?
Was Ab tested with avidin (to show that biotin is functional)? Any data in this regard?

Read More

Abcam community

Verified customer

Asked on May 09 2012

Answer

Thank you for contacting us.

The lab informed me that this antibody is tested against a RFP control proteinfrom Discosoma. We do not QC test the antibody against the DsRed variants you describe, but we have never receive a report of non-reactivity with dsRed variants. DsRed differs from our RFP by only a few amino acids and since our antibody is polyclonal against the whole RFP, it should detect any dsRed variant.

We use streptavidin HRP to test the function of the biotin.

Based on this information I have updated our datasheet to reflect this better and to be less misleading. I apologize about this, but am grateful that you pointed this out to us..

For our records, could you please send me a few more details about the staining that did not work (e.g. Antibody dilution, blocking conditions, sample type, detection systems used)? That would be of great help. Thank you.

In order to resolve this issue, I'd be happy to offer you a replacement, credit or refund if the antibody was purchased within the last 6 months. Please let me know your order or PO number in that regard as well as which option would be best for you..

I look forward to hear back from you
Please do not hesitate to contact me if you need any further advice or information.

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

Read More

Abcam Scientific Support

Answered on May 09 2012

Question

I was wondering whether your RFP antibody ab34771 also recognizes tdTomato? Thank you,

Read More

Abcam community

Verified customer

Asked on Apr 25 2012

Answer

Thank you for contacting us regarding ab34771.

Our anti RFP is designed to recognize all RFP variants from Discosoma, including TdTomato. While we do not test our lots of anti RFP against this variant, we have never had a complaint of non-reactivity of this product against any discosoma RFP variant.

This product is covered by our Abpromise guarantee. We provide scientific support, replacement or refund should this product not perform as indicated on the datasheet. More information on our Abpromise may be found at the following link:
https://www.abcam.com/index.html?pageconfig=abpromise

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.



Use our products? Submit an Abreview. Earn rewards!

https://www.abcam.com/abreviews

Read More

Abcam Scientific Support

Answered on Apr 25 2012

Question

We have paraffin embedded sections of mouse cartilage from mice with an mcherry reporter gene. Would Anti-RFP antibody (Biotin) (ab34771) react with our samples?

Read More

Abcam community

Verified customer

Asked on Feb 21 2012

Answer

Thank you for contacting Abcam regarding ab34771.


This Anti-RFP antibody is designed to recognize all variants of RFP, including mcherry, so the antibody will recognize this protein in your tissue samples.


I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

Read More

Abcam Scientific Support

Answered on Feb 21 2012

Question

Dear All,
a customer of ours is interested in AB34771 but in an unconjugated form.
I saw that you have different unconjugated anti RFP but the customer would like to know which is the correspondent unconjugated to AB34771. I hope my request is clear?
I'm looking for the same antidodies as AB34771(same immunogen) but not biotin-conjugated.

Thanks in advance for your help
Kind regards

Read More

Abcam community

Verified customer

Asked on Jan 17 2012

Answer

Thank you once again for your enquiries.

I am pleased to confirm we have added the unconjugated version of this antibody to the catalog for you. I can recommend to review the online datasheet for further details:

ab124754 Anti-RFP antibody
https://www.abcam.com/RFP-antibody-ab124754.html (or use the following: https://www.abcam.com/RFP-antibody-ab124754.html).

If you have any furtherquestions, please do not hesitate to contact me.

Read More

Abcam Scientific Support

Answered on Jan 17 2012

11-20 of 31 Abreviews or Q&A

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