Overview

  • Product name
  • Description
    Rabbit polyclonal to RFP
  • 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
  • Immunogen

    Synthetic peptide:

    VNGHEFEIEGEGEGR

    , corresponding to amino acids 22-36 of RFP from Discosoma sea anemone.

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, 1mg/ml BSA
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

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 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



  • Predicted band size : 26 kDa

    Western blot of recombinant RFP using 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

This product has been referenced in:
  • Lapierre LA  et al. Interaction of phosphorylated Rab11-FIP2 with Eps15 regulates apical junction composition. Mol Biol Cell 28:1088-1100 (2017). Read more (PubMed: 28228550) »
  • Gunzenhäuser J  et al. A Quantitative Approach to Evaluate the Impact of Fluorescent Labeling on Membrane-Bound HIV-Gag Assembly by Titration of Unlabeled Proteins. PLoS One 9:e115095 (2014). WB . Read more (PubMed: 25493438) »

See all 7 Publications for this product

Customer reviews and Q&As

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 ap...

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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
Username

Abcam user community

Verified customer

Submitted Dec 22 2015

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
Username

Abcam user community

Verified customer

Submitted Nov 09 2015

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, ...

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Thank you for your patience.

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

GAGAGAGAGAMSASSEDVITEFMRFKVKMEGTVNGHEFEIEGEGEGRPYEGHNTVKLKVT
KGGPLPFAWDILSPQFQYGS...

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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 wou...

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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 alw...

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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...

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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: http://www.abcam.com/index.html?datashe...

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Thank you very much for your interest in Abcam anti-RFP antibodies. ab28664 was recommended to your colleague because the homology between the immunogen and the mCherry is 100%. ab62341 was raised using the whole RFP, which has 82% homology with mCher...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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