Anti-RFP antibody (ab28664)

Reviews (4)Q&A (16)References (12)
Western blot - Anti-RFP antibody (ab28664)
  • Immunocytochemistry/ Immunofluorescence - Anti-RFP antibody (ab28664)

Key features and details

  • Rabbit polyclonal to RFP
  • Suitable for: WB, ICC/IF
  • Reacts with: Human, Recombinant fragment
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-RFP antibody [EPR18992] (ab185921)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-RFP antibody
    See all RFP primary antibodies

  • Description

    Rabbit polyclonal to RFP

  • Host species

    Rabbit

  • Specificity

    ab28664 detects RFP tagged proteins. This antibody is specific for the RFP tag. It shows very low reactivity to GFP or GFP tagged proteins. ab28664 non-specifically detects a 45 kDa protein from E. coli extracts. This non-specific cross-reactivity is not present in mammalian samples.

  • Tested applications

    Suitable for: WB, ICC/IFmore details

  • Species reactivity

    Reacts with: Human, Recombinant fragment

  • Immunogen

    Synthetic peptide:

    VNGHEFEIEGEGEGR

    , corresponding to amino acids 22-36 of RFP from Discosoma sea anemone.
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • General notes

    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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.

  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituents: PBS, 0.1% BSA
  • Concentration information loading...

  • Purity

    Immunogen affinity purified

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab28664 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 (3)
1/1000. Predicted molecular weight: 26 kDa.

Use at a concentration of 2 µg/ml. Block membrane in 5% milk and incubate primary and secondary antibodies in milk.
ICC/IF (1)
1/50 - 1/200.
Notes
WB
1/1000. Predicted molecular weight: 26 kDa.

Use at a concentration of 2 µg/ml. Block membrane in 5% milk and incubate primary and secondary antibodies in milk.
ICC/IF
1/50 - 1/200.

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 - Anti-RFP antibody (ab28664)
    Western blot - Anti-RFP antibody (ab28664)

    Western blot of recombinant RFP using ab28664.

  • Immunocytochemistry/ Immunofluorescence - Anti-RFP antibody (ab28664)
    Immunocytochemistry/ Immunofluorescence - Anti-RFP antibody (ab28664)

    Immunofluorescent analysis of recombinant Red Fluorescent Protein (RFP)-transfected HeLa cells. Cells were transfected with a pCMV RFP C-Myc plasmid and 48 hours post-transfection cells were fixed with formalin and permeabilized with 0.1% Triton X-100 for 15 minutes at room temperature. Cells were blocked with 3% BSA for 15 minutes at room temperature, and then probed with ab28664 at a dilution of 1/50 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight® 488-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1/400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.

References (12)

Publishing research using ab28664? Please let us know so that we can cite the reference in this datasheet.

ab28664 has been referenced in 12 publications.

  • Schwirz J  et al. Bicistronic expression and differential localization of proteins in insect cells and Drosophila suzukii using picornaviral 2A peptides. Insect Biochem Mol Biol 119:103324 (2020). PubMed: 31978587
  • Chang X  et al. PADI3 induces cell cycle arrest via the Sirt2/AKT/p21 pathway and acts as a tumor suppressor gene in colon cancer. Cancer Biol Med 16:729-742 (2019). PubMed: 31908891
  • Chai Z  et al. PADI3 plays an antitumor role via the Hsp90/CKS1 pathway in colon cancer. Cancer Cell Int 19:277 (2019). PubMed: 31708688
  • Nakagawa N  et al. APC sets the Wnt tone necessary for cerebral cortical progenitor development. Genes Dev 31:1679-1692 (2017). PubMed: 28916710
  • Lapierre LA  et al. Interaction of phosphorylated Rab11-FIP2 with Eps15 regulates apical junction composition. Mol Biol Cell 28:1088-1100 (2017). PubMed: 28228550
View all Publications for this product

Customer reviews and Q&As

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1-10 of 20 Abreviews or Q&A

Western blot abreview for Anti-RFP antibody

 Inconclusive
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (human induced pluripotent stem cells)
Gel Running Conditions
Reduced Denaturing (4~12% NuPAGE Bris-Tris Gel)
Loading amount
20 µg
Treatment
mApple-tagged transgene
Specification
human induced pluripotent stem cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Feb 24 2021

Immunocytochemistry/ Immunofluorescence abreview for Anti-RFP antibody

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEK293T)
Permeabilization
Yes - 0.3% Triton X-100
Specification
HEK293T
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Dec 22 2015

Western blot abreview for Anti-RFP antibody

 Average
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (mApple-CD81 transfected in CHO cells)
Gel Running Conditions
Reduced Denaturing (5-20% gradient gel)
Loading amount
50 µg
Specification
mApple-CD81 transfected in CHO cells
Blocking step
Milk as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Read More

Abcam user community

Verified customer

Submitted Nov 09 2015

Question

In cultured transiently transfected 293T cells there is a small amount of background with the anti-RFP antibody after 1-2 hrs exposure but no bands could be detected at the correct size for the ARF-RFP fusion (˜40kDa. See attached: lanes 1-3 should have expression, with lane 2 having the most protein) . The blot was then stripped and probed with anti-p19 and a band of the correct size could be detected and was induced to even higher degree in the presence of rtTA and following the addition of doxycycline (lane 2; ARF-RFP is driven from a doxycycline- and rtTA-dependent TRE promoter). Thus I am confident that the construct works and the protein is generated in vitro. If used on transgenic mouse tissue samples, there is more background (as observed with many antibodies) but no band corresponding to ARF-RFP.



The lysates were boiled at 95’C for 5 minutes.



Yes, our secondary antibody works (checked against other anti-Rabbit antibodies in the lab).



I cannot find the purchase order number right now but have emailed our accounts team and will get back to you.



I have indeed checked expression under a fluorescent microscope and I see very robust expression in cells transfected with the vector.



I hope some of that info helps. Do not hesitate to contact me if you require more information.



Many thanks,

Read More
Answer

Thank you for your enquiry regarding ab28664 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means that in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

The protocol looks absolutely fine to me. I could offer you either a new vial of ab28664 or an alternative antibody against the same target or a credit note which you can use for future purchase.





Please dolet me know how you wish to proceed with your enquiry. . I look forward to hearing from you soon.

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Question

Hi, I was wondering if this product will detect mCherry? We have designed a doxycycline inducible stable cell line in MDCK cells. We use mCherry as one of our controls and need an antibody that will detect it in Western blots. If this antibody is able to detect mCherry, may I also get a quote on this item.

Read More
Answer

Thank you for contacting us.

To our knowledge, the antibody ab62341 has not been tested for reactivity with mCherry. I suggest trying ab28664, if you only plan to use it for western blotting, because the immunogen sequence, VNGHEFEIEGEGEGR, is 100% conserved in the mCherry sequence at the following URL.

http://www.ncbi.nlm.nih.gov/protein/114794217

This antibody has not been tested for reactivity with mCherry either but we expect it has a higher likelihood of cross-reaction than ab62341, given that the immunogen for ab62341 is full-length RFP (http://www.uniprot.org/uniprot/Q9U6Y8.fasta), which has only 85% identity with full-length mCherry.

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

Click here (or use the following: https://www.abcam.com/RFP-antibody-ab28664.html).

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Question

Here is the sequence. It would be great if you can check it.

Redstar2 Tag


GGAGCTGGAGCTGGTGCAGGTGCTGGTGCAATGAGTGCTTCTTCTGAAGATGTCATCACTGAATTCATGAGATTCAAGGTTAAAATGGAAGGTACTGTTAACGGCCACGAATTCGAAATCGAAGGTGAAGGTGAGGGTAGACCATATGAAGGTCACAACACAGTCAAGTTGAAGGTTACTAAGGGTGGTCCACTGCCATTCGCTTGGGACATCTTGTCTCCACAATTCCAATACGGTTCTAAGGTCTACGTCAAGCACCCAGCTGACATTCCAGACTACAAGAAGTTGTCCTTCCCAGAAGGTTTCAAGTGGGAAAGGATCATGAACTTCGAAGACGGTGGCGTTGTTACTGTTACTCAAGACTCCTCCTTGCAAGACGGTTGTTTCATCTACAAGGTCAAGCTCATTGGTGTCAACTTCCCATCTGACGGTCCAGTCATGCAAAAGAAGACTATGGGTTGGGAAGCTTCTACCGAACGTTTGTACCCAAGAGACGGTGTCTTGAAGGGTGAAATCCACAAGGCCTTGAAGTTGAAGGACGGTGGTCACTACTTGGTCGAATTCAAGTCTATCTACAAGGCCAAGAAGCAAGTCCAATTGCCAGGCTATTACTACGTTGACTCTAAGTTGGACATCATCTCTCACAACGAAGACTACACTATCGTCGAACAATACGAACGTACTGAAGGTAGACACCACTTGTTCTTGTAA

Read More
Answer

Thank you for your patience.

Using the Expasy translate tool, I converted the nucleotide sequence you provided to the following protein sequence:

GAGAGAGAGAMSASSEDVITEFMRFKVKMEGTVNGHEFEIEGEGEGRPYEGHNTVKLKVT
KGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERIMNFEDGGVVTVTQ
DSSLQDGCFIYKVKLIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHKALKLKD
GGHYLVEFKSIYKAKKQVQLPGYYYVDSKLDIISHNEDYTIVEQYERTEGRHHLFL

Three of our RFP antibodies were raised against immunogen sequences that are 100% conserved in Redstar2: ab28664, ab40740, and ab41336. While we have not tested these antibodies for detecting Redstar2, it is very likely that they would be suitable for that purpose.

I hope this helps, please let me know if you need any additional information or assistance.

Read More

Question

We are looking for an anti-RFP antibody that will recognise a mutated form of RFP. We would like to check if ab28664 antibody works with RedStar2, made from a combination of T4-DsRed and the RedStar mutant ( Janke et al, Yeast 2004;21: 947-962). Do you know if this antibody works for Western Blotting on RedStar2 or could we have a small quantity to test.

Read More
Answer

Thank you for contacting us.

We have not validated this antibody for use with RedStar2. If you could provide the amino acid sequence for this protein I will be happy to check the homology with our RFP antibodies to see if we have one that would be likely to cross-react.

Unfortunately, we do not offer free or trial sized samples of any of our products. I am sorry I could not be more helpful, please let me know if you need any additional information or assistance.

Read More

Question

Will this antibody recognize the monomeric form of RFP?

Read More
Answer

Thank you for your inquiry.


The antibody was raised against amino acids 22-36 of the RFP protein, which is made up of a single 231 amino acid polypeptide chain. The lab said that theprotein doesn't exist in any other form,that it is always a monomer.


I hope this information helps. Please contact us with any other questions.

Read More

Question

we wish to purchase the ab28664 antibody. Please send us the discount code fortesting it in ICC/IF and submitting feedback.

Thanks

Read More
Answer

I am very pleased to hear you would like to accept our offer and test ab28664 in ICC/IF. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for ICC/IF and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. 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.

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Question

Hi, I am writing in to ask about the suitability of RFP antibodies on your catalog to probe for a mCherry-tagged protein on WB. Will there be any binding issues? And for EGFP-tagged proteins, are all GFP antibodies able to detect the EGFP on WB? Many thanks.

Read More
Answer

Thank you for your enquiry. Indeed, we do have a few RFP antibodies available that also recognise mCherry and are suitable for Western blot, as stated on each datasheet: ab28664: Click here (or use the following: https://www.abcam.com/RFP-antibody-ab28664.html). ab105750: Click here (or use the following: https://www.abcam.com/RFP-antibody-ab105750.html). ab109809: Click here (or use the following: https://www.abcam.com/RFP-antibody-RFPH8-ab109809.html). To our knowledge, ab28664 has not been tested in specifically tested for cross-reactivity with mCherry yet. However, the immunogen shares a 100% sequence similarity with mCherry and so it is very likely that it will work. Furthermore, we do not currently have an image of ab105750 or ab109809 showing the detection of mCherry. Therefore, I can offer a discount off a future purchase if you buy one of these antibodies now, test it in Western blot with mCherry and submit feedback to us in the form of an Abreview (including an image). 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 which antibody you would like to test. I will then send a discount code. This code must be issued before purchasing the antibody so please wait for my reply before ordering. 2. Purchase the antibody either by phone, fax, or online (www.abcam.com). 3. Test it in WB with mCherry. 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 more about our Abreview system, 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. Regarding the detection of EGFP proteins, I am happy to let you know that most of our GFP antibodies also detect the enhanced GFP. This is also mentioned on each individual datasheet, such as for ab290 (Click here (or use the following: https://www.abcam.com/GFP-antibody-ab290.html).). All of the information we have for each of our products is listed on the online datasheet for your convenience. These are updated as soon as any new information is brought to our attention. Please be reassured that all our products are covered by our Abpromise: We will happily offer Scientific Support in the event that a product is not functioning in the applications and species cited on the product datasheet (and the problem has been reported within 6 months of purchase). If it appears that the antibody is faulty, a replacement, credit or refund will be offered (https://www.abcam.com/Abpromise). 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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