Overview

  • Product name
    Anti-V5 tag antibody (Agarose)
    See all V5 tag primary antibodies
  • Description
    Goat polyclonal to V5 tag (Agarose)
  • Host species
    Goat
  • Conjugation
    Agarose
  • Tested applications
    Suitable for: IPmore details
  • Immunogen

    Full length native

    GKPIPNPLLGLDST

    (V5) (purified): conjugated to KLH.

  • General notes
    Provided as 400µl of slurry (200µl gel + 200µl buffer). 100µg of the affinity purified antibody is attached to the gel at a ratio of 500µg antibody/ml of gel or 250µg IgG/ml of 50% slurry.

    Affinity purified antibodies were coupled to agarose beads using a cyanogen bromide method.

Properties

Applications

Our Abpromise guarantee covers the use of ab1229 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
IP Use at an assay dependent concentration. Use at a concentration of 15 - 20 µl of gel slurry per 0.1 to 1 mg of protein lysate or extract.

Target

  • Relevance
    The V5 epitope tag is derived from a small epitope (Pk) present on the P and V proteins of the paramyxovirus of simian virus 5 (SV5). The V5 tag is usually used with all 14 amino acids (GKPIPNPLLGLDST), although it has also been used with a shorter 9 amino acid sequence (IPNPLLGLD).
  • Alternative names
    • GKPIPNPLLGLDST epitope tag antibody
    • GKPIPNPLLGLDST tag antibody
    • Protein Rev antibody
    • Regulator of expression of viral proteins antibody
    • rev antibody
    • V5 epitope tag antibody
    see all

References

This product has been referenced in:
  • Liu T  et al. Glycosylation controls sodium-calcium exchanger 3 sub-cellular localization during cell cycle. Eur J Cell Biol 97:190-203 (2018). Read more (PubMed: 29526322) »
  • Yalla K  et al. FBXW7 regulates DISC1 stability via the ubiquitin-proteosome system. Mol Psychiatry 23:1278-1286 (2018). Read more (PubMed: 28727686) »
See all 13 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Question
Answer

J’ai contacté le laboratoire et un tampon de RIPA pourrait peut-être fonctionner mais nous ne pouvons pas le confirmer car nous ne l’avons pas testé.

J’aimerais cependant mentionner qu’en utilisant un détergent assez fort comme du SDS il y a plus de risque de déstabilisation du complexe anticorps-antigène, ce qui pourrait diminuer l’intensité du signale. Si ceci est le cas, je vous recommande de suivre notre protocole standard pour l’IP.

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Application
Immunoprecipitation
Immuno-precipitation step
Other - ab1229
Sample
Human Cell lysate - whole cell (HEK293T)
Specification
HEK293T
Treatment
transfected with plasmid DNA
Total protein in input
5e+006 cells

Ms. Christina Engel

Verified customer

Submitted Oct 10 2013

Answer



Our labs have tested this product using 0.1 % triton X-100.

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Answer

Thank you for contacting us.
Please find below the purification procedure using antibodies on agarose beads:
1. The agarose beads with the antibody bound to them are ready for use when you receive them.
2. Place the beads in a column and equilibrate with the appropriate buffer, usually two to three column volumes of PBS or TBS (1M NaCl).
3. Pass the media or serum that contains the protein of interest through the column.
4. Collect the “flow through” of unbound protein.
5. Wash the column with equilibrating buffer until the A280 = 0.01
7. Elute the desired protein with one of several buffers of your choice that is compatible with the activity of your protein: A. An acid buffer down to pH of 2.5*, i.e. 5% acetic acid or glycine HCl. B. 3 or 4 M MgCl2 (magnesium chloride). C. 6 or 8 M Urea. D. 4 or 6 M Guanidine. E. 3 to 4 M Potassium Isothiocyantate. F. elution with peptide**
8. Regenerate the column by passing several volumes of elution buffer over column/beads (e.g. with three column volumes of glycine-HCI, pH 2.5***) Immediately re-equilibrate with equilibrating buffer (PBS or TBS) until the effluent is a neutral pH.or dialyze against a desired buffer.
9. Store the agarose-antibody beads in a buffer containing a preservative i.e. 0.1% sodium azide at 2-8 deg C.. They may be used many times.
* Occasionally, low pH may cause the eluted protein to aggregate. In such cases choose an alternative buffer for elution, for example 3 M sodium thiocyanate. The column may lose activity after prolonged exposure to low pH.
** Elution by Peptide: This is a milder elution method. Elute the bound tagged fusion protein by adding 5 X 1 column volume aliquots of a solution containing 100 mg/ml peptide in PBS. Note: peptide has a detectable absorbency at280 nm and also interferes in other protein determination assays that are based on peptide bonds. Therefore, it is recommended to determine the eluted amount by Coomassie staining of SDSPAGE relative to a known standard.
***Do not leave the column in glycine-HCl for longer than 20 minutes.
I hope it is helpful for you. If not, please do not hesitate to contact us to assist you further.

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Answer

Thank you for your enquiry. The buffer that we know it works in has a concentration of 0.1% SDS and 0.5% deoxycholate. I hope this information will be helpful. If there is anything else that I can help you with, please do not hesitate to contact me.

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Answer

Thank you for your enquiry. Theoretically this will work, but to the best of our knowledge it has not been proven to work satisfactorally. I hope this information helps. Please do not hesitate to contact us if you need anything further.

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Answer

Thank you for your enquiry. Following immunoprecipitation, some heavy and light IgG chains may be eluted with reducing buffers such as Laemmli. This may cause interference with your V5-tagged protein. The best and cleanest approach would be to IP using a V5 antibody raised in one species (e.g. mouse) and perform a western blot using an antibody raised in another (e.g. rabbit). I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for your enquiry. Tris based buffers will not have an appreciable influence on the agarose immobilized antibody. Some heavy and light IgG chains will be eluted with reducing buffers such as Laemmli.

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Answer

We acknowledge receipt and would like to thank you for your request of bulk quote for the ab1229. I am currently working on our best quotation and will be contacting you in the next couple of days.

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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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