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  1. Link

    agarose-v5-tag-antibody-ab1229.pdf

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Tags & Cell Markers Epitope Tags V5 Tag
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Agarose Anti-V5 tag antibody (ab1229)

  • Datasheet
  • SDS
Reviews (1)Q&A (8)References (17)

Product price, shipping and contact information

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Promotion Information

Abpromise

Guaranteed product quality, expert customer support.

Find out more.

Key features and details

  • Agarose Goat polyclonal to V5 tag
  • Suitable for: IP
  • Reacts with: Species independent
  • Conjugation: Agarose
  • Isotype: IgG

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Overview

  • Product name

    Agarose Anti-V5 tag antibody
    See all V5 tag primary antibodies
  • Description

    Agarose Goat polyclonal to V5 tag
  • Host species

    Goat
  • Conjugation

    Agarose
  • Tested applications

    Suitable for: IPmore details
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Full length native

    GKPIPNPLLGLDST

    (V5) (purified): conjugated to KLH.
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 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.

    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.
  • Storage buffer

    pH: 6.8
    Preservative: 0.1% Sodium azide
    Constituents: 0.0268% PBS, 0.58% Sodium chloride
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Epitope Tags
    • V5 Tag
    • Tags & Cell Markers
    • Epitope Tags
    • Agarose Conjugates
    • Signal Transduction
    • Antibodies
    • v5 tag

Associated products

  • Immunizing Peptide (Blocking)

    • V5 tag peptide (ab15829)
  • Isotype control

    • Agarose Goat IgG, polyclonal - Isotype Control (ab104155)

Applications

The Abpromise guarantee

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 (1)
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.
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

Protocols

  • Immunoprecipitation protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (17)

Publishing research using ab1229? Please let us know so that we can cite the reference in this datasheet.

ab1229 has been referenced in 17 publications.

  • Zhou YT  et al. A Species-Correlated Transitional Residue D132 on Human FMRP Plays a Role in Nuclear Localization via an RNA-Dependent Interaction With PABP1. Neuroscience 404:282-296 (2019). PubMed: 30742966
  • Cockman EM  et al. Identification of the Selenoprotein S Positive UGA Recoding (SPUR) element and its position-dependent activity. RNA Biol 16:1682-1696 (2019). PubMed: 31432740
  • Greenberg RS  et al. Single Amino Acid Change Underlies Distinct Roles of H2A.Z Subtypes in Human Syndrome. Cell 178:1421-1436.e24 (2019). PubMed: 31491386
  • Pickar-Oliver A  et al. Targeted transcriptional modulation with type I CRISPR-Cas systems in human cells. Nat Biotechnol 37:1493-1501 (2019). PubMed: 31548729
  • Liu T  et al. Glycosylation controls sodium-calcium exchanger 3 sub-cellular localization during cell cycle. Eur J Cell Biol 97:190-203 (2018). PubMed: 29526322
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-9 of 9 Abreviews or Q&A

Immunoprecipitation abreview for Anti-V5 tag antibody (Agarose)

Excellent
Abreviews
Abreviews
abreview image
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
Read More

Ms. Christina Engel

Verified customer

Submitted Oct 10 2013

Question

Est-ce que je peux remplacer votre cell lysis buffer par un tampon RIPA pour l'IP ?

Read More

Abcam community

Verified customer

Asked on Dec 23 2014

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.

Read More

Elisa Thomas

Abcam Scientific Support

Answered on Dec 23 2014

Question

What is the concentration of detergents that can be used to perform the pull down assay with this product

Read More

Abcam community

Verified customer

Asked on Feb 25 2013

Answer



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

Read More

Abcam Scientific Support

Answered on Feb 25 2013

Question

Hi, I ordered the Anti-V5 tag antibody to concentrate my V5-tagged protein.
Do you have a protocol that for how long should the agarose beads be incubated with my solution of interest and in which conditions? Do you recommend certain kind of centrifuging afterwards, for example 30 min 13000 rpm?
Is it possible to dissociate the protein of interest from the beads afterwards?
with best regards

Read More

Abcam community

Verified customer

Asked on Mar 05 2012

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.

Read More

Abcam Scientific Support

Answered on Mar 05 2012

Question

does the ab1229 work in buffer containg SDS and/or deoxycholate, if yes what are the maximum concentrations? thanks

Read More

Abcam community

Verified customer

Asked on Jun 03 2008

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.

Read More

Abcam Scientific Support

Answered on Jun 03 2008

Question

I am trying to purify protein using this product. Can the protein be competed off the beads with V5 peptide instead of boiled off?

Read More

Abcam community

Verified customer

Asked on Dec 19 2005

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.

Read More

Abcam Scientific Support

Answered on Dec 22 2005

Question

I would like information about your agarose-immobilized anti-V5 antibody. The intended application is immunoprecipitation followed by western blot. My V5-fusion protein is ~55kb, so I am concerned about interference with the IgG heavy chain. Will the antibody remain bound to the agarose after I boil my samples in reducing SDS loading buffer (containing DTT)?

Read More

Abcam community

Verified customer

Asked on Dec 14 2005

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.

Read More

Abcam Scientific Support

Answered on Dec 15 2005

Question

I am interested in using your agarose-conjugated anti-V5 antibody for immunoprecipitation, but I am concerned about the stability of the cyanogen bromide linkage. I have read, for example, that CnBr linkages may be incompatible with Tris-based buffers. Have you found this to be the case with your crosslinked antibodies? Also, will the linkage be disrupted by boiling in Laemmli buffer?

Read More

Abcam community

Verified customer

Asked on Jul 20 2004

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.

Read More

Abcam Scientific Support

Answered on Jul 27 2004

Question

I messed up the story I apologize, I wanted to order the same amount that I ordered last time (if possible for the same price).

Read More

Abcam community

Verified customer

Asked on Oct 08 2003

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.

Read More

Abcam Scientific Support

Answered on Oct 13 2003

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