Overview

  • Product name
    Anti-Phosphoserine antibody [PSR-45]
    See all Phosphoserine primary antibodies
  • Description
    Mouse monoclonal [PSR-45] to Phosphoserine
  • Host species
    Mouse
  • Tested applications
    Suitable for: Dot blot, IP, Indirect ELISA, WBmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    Other Immunogen Type corresponding to Phosphoserine conjugated to keyhole limpet haemocyanin.
    Sequence:

    phospho-serine


    (Peptide available as ab100812)

  • Positive control
    • WB: rat brain cortex lysate ( a main 50kDa band is seen by detection with biotin-avidin AEC staining) ELISA: phosphoserine conjugated to BSA
  • General notes

    Abcam is committed to meeting high standards of ethical manufacturing and has decided to discontinue this product by June 2019 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We would recommend antibody ab9332 as a replacement.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab6639 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
Dot blot Use at an assay dependent concentration.
IP 1/2000.
Indirect ELISA 1/4000.
WB 1/500 - 1/1000. Do not use milk as a blocking agent or in diluents, as milk casein is phosphorylated at several serine residues. BSA is recommended instead. Stronger blocking agent percentage is recommended (e.g. 10%), in addition to varying loading amounts and shorter incubation time (1-2h RT)

Target

  • Relevance
    Changes in the serine/threonine phosphorylation state of a protein in response to various external stimuli can have profound effects on cellular signal transduction, apoptosis and carcinogenesis. The reagents, including phosphorylated protein/peptides, antibodies against the phosphospecific amino acid, are important tools to explore the activation of serine, threonine or tyrosine containing proteins. An aberrant protein phosphorylation is a hallmark of human disease, and the enzymes, particularly protein kinases, which control protein phosphorylation are recognized as a major new drug target family.
  • Alternative names
    • phospho-Ser antibody
    • pS antibody
    • pSER antibody

Images

  • Anti-Phosphoserine antibody [PSR-45] (ab6639) at 1/200 dilution + Drosophila melanogaster whole tissue lysate 40 ug per lane at 40 µg

    Secondary
    Sheep anti mouse (HRP) at 1/2000 dilution

    Performed under reducing conditions.


    This image is an edited version of an image submitted courtesy of an Abreview submitted on 17 August 2005. We do not have any further information relating to this image.

    See Abreview

  • ab6639 used in Immunoprecipitation.
    Whole cell lysate prepared from murine mammary cells was loaded at 250µg.
    Cells were treated with 5ng/ml TGF beta for varying time periods.
    Protein A was used for the Immunoprecipitation step.
    ab6639 used at a 1/2000 dilution for 10 hours at 4°C.
    Western blot was performed with an HRP anti-PCBP1 polyclonal antibody, 1/10000 dilution.

    See Abreview

References

This product has been referenced in:
  • Nyati KK  et al. TLR4-induced NF-?B and MAPK signaling regulate the IL-6 mRNA stabilizing protein Arid5a. Nucleic Acids Res 45:2687-2703 (2017). WB . Read more (PubMed: 28168301) »
  • Liu H  et al. Interleukin-4 and interleukin-13 increase NADPH oxidase 1-related proliferation of human colon cancer cells. Oncotarget 8:38113-38135 (2017). Read more (PubMed: 28498822) »
See all 17 Publications for this product

Customer reviews and Q&As

1-10 of 30 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (NSCLC)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
30 µg
Treatment
0-100 nM volasertib for 2 Hrs
Specification
NSCLC
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 11 2017

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number ***.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Answer

Thank you for your reply.
I am sorry to hear that the suggestions we discussed did not improve the results.

Yes, I'd be happy to replace, credit or refund the antibodies. Please just let me know which option is best for you. In case of a replacement, please let me know which antibodies you would like to receive free of charge in return.

I look forward to hear back from you and resolve this case for you.

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Answer

Thank you for your emails and the additional details you sent.

Yes, the use of PBS is a good point. In case of using anti-phospho antibodies this can actually cause high background due to the presence of phosphate in the buffer. I would suggest using rather TBS instead.
Right now, we do not know if the problem stems from the PBS alone or also fromthe antibody. Thus, I would suggest trying once or twice with TBS or TBST. Shouldthe backgroundnot become lower, then theantibody probably contributes to the background as well.

Would you be able to try this out and then let me know?

Also, I wanted to let you know that via Epitomics (an Abcam company) we are now offering custom antibody service, thus if you would need a specific anti-phospho-MAL2 antibody, they might be able to produce one. Please check the following links:

https://www.abcam.com/epitomics
https://www.abcam.com/index.html?pageconfig=resource&rid=15281
http://www.epitomics.com/services


I look forward to hear back from you.

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https://www.abcam.com/abreviews

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Answer

Thank you for contacting us.

I am sorry to hear that the 2 antibodies have not been working as expected.

Your protocol looks fine in general. However, both antibodies are directed against ANY phospho-site on a serine or threonine of any protein, thus they will detect all proteins that have these modifications. If you are looking for a specific protein of interest, it would be difficult to determine if you are picking up this protein or not by using a general phospho-Ser/Thr antibody.

Where would you expect the specific protein band to be detected (which MW)? You would need to compare your blots with one for the total protein of your interest, or even cut your blot lengthswise and do the blotting side by side.

Also, if you are detecting phospho-sites, you would need to add protease as well as phosphatase inhibitors when preparing your lysates. Otherwise, most of the phosphogroups might have been removed by the phosphatases when the cells are lysed. I am not sure which phosphatase is being inhibited by okadaic acid, butI would suggest using alsoother phospatase inhibitors (e.g. sodium vanadate) as certain inhibitors inhibit only certain phosphatases.

What residues are phosphorylated on your protein of interest (Ser, Thr, Tyr)? Under which conditions is your protein MAL2 phosphorylated? Maybe phosphorylation will need to be induced by a specific treatment or induction?

Some of your blots show black backgrounds or are completely black with inverted (white) bands. This is often an indication that either the ECL reagents could be contaminated or that too much antibody was used or the exposure time is too long.

Have you tried a no primary control (only secondary and noprimary antibody)? This would help to determine if any of the heavy backgroundstems from the secondary antibody alone, as well as if there is any cross reactivity of the secondary antibody withtheFCS or bovine calf serum used for blocking. Theoretically there should be no cross-reactivity, but practically this is not always the case. BSA would be the better blocking reagent.

Also, please let me know howyou were preparing your samples in detail and how much protein was loaded per lane.

I hope these suggestions are helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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https://www.abcam.com/abreviews

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Question
Answer

Thank you for your telephone enquiry last week.

I am sorry to confirm that ab6639 Phosphoserine antibody [PSR-45] has only been tested in rat brain tissue bythe originator, and so we have no images for human samples.

There have, however, been several publications citing the use of this antibody with human samples:

Barattini, P., et al., Expression and DNA binding activity of the Ku heterodimer in bladder carcinoma Cancer 92, 2484-2492, (2001)

Huang, H., et al., A Protein Kinase Activity Associated with Epstein-Barr Virus BGLF4 Phosphorylates the Viral Early Antigen EA-D in vitro J. Virol. 74, 3093-3104, (2000)

Gershburg, E., et al., Hyperphosphorylation of EBNA2 by Epstein-Barr Virus Protein Kinase Suppresses Transactivation of the LMP1 Promoter J. Virol. 79, 5880-5885, (2005)

Bruzzone, S., et al., Abscisic Acid Is An Endogenous Stimulator Of Insulin Release From Human Pancreatic Islets With Cyclic ADP Ribose As Second Messenger. J. Thorac. Cardiovasc. Surg. 283, 32188-97, (2008)

Barber, M.A., et al., The guanine-nucleotide-exchange factor P-Rex1 is activated by protein phosphatase 1a. Biochem. J. 443, 173-83, (2012)

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

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Answer

Thank you for contacting us.

To our knowledge this product has not been tested in IHC. For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:
The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

Please let me know if you would like to participate in this program. Please do not hesitate to contact us if you need any more advice or information.



Use our products? Submit an Abreview. Earn rewards!

https://www.abcam.com/abreviews

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Answer

Thank you for your reply and providing that information.

Your credit note ID is ********.

I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

Please let me know if there is anything else I can help you with.

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Answer

Thank you for contacting Abcam.

Based on the information that you have provided, I do not think that there is any protocol tips that would help resolve the issue, as I think you have tried everything.

Under our Abpromise, which is good for 6 months, we guarantee that this antibody will work in western blot and since it is not, I will be happy to either replace or refund the cost of this antibody. If you could please provide me with the original order details for this antibody and the catalogue number of the product you would like as a replacement, I would be happy to help resolve this issue for you.

Looking forward to your reply.

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Answer

Merci pour votre email.

J'ai demandé à notre service comptabilité de mettre en place un avoir référence CN.

Votre référence CN est ******.

Voici les manières dont vous pouvez utiliser cette référence CN:
(1) la faire valoir contre la facture originale si celle-ci n'as pas encore été payée
(2) la faire valoir contre une prochaine facture
(3) obtenir un remboursement de la somme entière si vous n'avez pas de factures actuellement en cours avec Abcam.

Si vous souhaitez recevoir un remboursement au lieu d'un avoir, veuillez demander à votre service de gestion de prendre contact avec notre département comptabilité afin que nous puissions recueillir les informations nécessaires pour cette restitution. Notre service comptabilité peut être contacté par courriel à l'adresse mailto:creditcontrol@abcam.com ou par téléphone en utilisant le lien « Contactez-nous » dans le coin en haut à droite de notre site internet. Veuillez communiquer votre numéro d'avoir lorsque vous contactez notre service comptabilité.

Vous recevrez également la référence complète de la note de crédit par courriel ou par voie postale; cette référence commençant par les lettres CGB.

J'espère que cette expérience ne vous empêchera pas de commander à nouveau l'un de nos produits.

L'équipe technique reste à votre disposition pour plus de conseils.

Bonne continuation dans votre recherche.

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1-10 of 30 Abreviews or Q&A

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