Anti-Phospho - (Ser/Thr) Phe antibody (ab17464)

Reviews (5)Q&A (14)References (33)

Overview

  • Product name
    Anti-Phospho - (Ser/Thr) Phe antibody
  • Description
    Rabbit polyclonal to Phospho - (Ser/Thr) Phe
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, ELISA, IP, WBmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    Synthetic peptide corresponding to Phospho - (Ser/Thr) Phe . Immunogen = Synthetic phospho-serine/threonine-phenylalanine containing peptide.

  • Positive control
    • WB: Calyculin A treated A431 cells; Human THP-1 Whole Cell Extract; Mouse Total Lung Extract.
  • General notes

    The immunogen used is specific phosphorylated serine or threonine amino acids surrounded by phenylalanine. (Y/F/W)(S*/T*)F

Properties

Applications

Our Abpromise guarantee covers the use of ab17464 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
ICC/IF 1/100.
ELISA 1/1000.
IP 1/100.
WB 1/1000.

Block with 5% nonfat milk, 0.1% Tween-20. Dilute primary antibody in 5% BSA, 0.1% Tween-20.

Images

  • Western blot - Anti-Phospho - (Ser/Thr) Phe antibody (ab17464)
    Western blot - Anti-Phospho - (Ser/Thr) Phe antibody (ab17464)Image from Gao X et al, PLoS Pathog 5:e1000708 (2009), Fig S3.
    To determine if NleH could phosphorylate RPS3, we conducted in vitro kinase assays with NleH and RPR3 and analyzed the results by Immunoblotting with ab17464, phospho-Ser/Thr-specific antibody, following separation by SDS-PAGE.
  • Western blot - Anti-Phospho - (Ser/Thr) Phe antibody (ab17464)
    Western blot - Anti-Phospho - (Ser/Thr) Phe antibody (ab17464)This image is courtesy of an anonymous Abreview
    All lanes : Anti-Phospho - (Ser/Thr) Phe antibody (ab17464) at 1/1000 dilution

    Lane 1 : Human THP-1 Whole Cell Extract
    Lane 2 : Mouse Total Lung Extract

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Alkaline phosphatase conjugated goat anti-rabbit antibody

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 5 minutes

    See Abreview

  • Western blot - Anti-Phospho - (Ser/Thr) Phe antibody (ab17464)
    Western blot - Anti-Phospho - (Ser/Thr) Phe antibody (ab17464)
    All lanes : Anti-Phospho - (Ser/Thr) Phe antibody (ab17464) at 1/1000 dilution

    All lanes : calyculin A treated A431 cells

References

This product has been referenced in:
  • Wei X  et al. Targeting the Dvl-1/ß-arrestin2/JNK3 interaction disrupts Wnt5a-JNK3 signaling and protects hippocampal CA1 neurons during cerebral ischemia reperfusion. Neuropharmacology 135:11-21 (2018). IP ; Rat . Read more (PubMed: 29510185) »
  • Po A  et al. Noncanonical GLI1 signaling promotes stemness features and in vivo growth in lung adenocarcinoma. Oncogene 36:4641-4652 (2017). Read more (PubMed: 28368412) »
See all 33 Publications for this product

Publishing research using ab17464? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 19 Abreviews or Q&A

Western blot abreview for Anti-Phosphoserine/threonine antibody

 Good
Abreviews
abreview image
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (hepatocytes)
Specification
hepatocytes
Treatment
20 ug/ml poly(I:C)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More

Abcam user community

Verified customer

Submitted Jun 26 2014

Western blot abreview for Anti-Phosphoserine/threonine antibody

 Below Average
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Loading amount
100 µg
Specification
HeLa
Treatment
UV irradiation
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More

Abcam user community

Verified customer

Submitted May 13 2013

Question

We wanted to check the general phosphorylated serine/threonine level of our protein (especially for phospho-threonine). So we would like to have a antibody that could specific recognize all phosphorylated serine/threonine residues (even p-Tyr), but not highly dependent on their neighboring residues. I found three different types of antibodies might do the job:

Anti-Phosphoserine/threonine/tyrosine antibody [SPM101] (ab15556)

Anti-Phosphoserine/threonine antibody (ab17464)

Anti-Phosphothreonine antibody (ab9337)

Any suggestion for choosing a good one from these three for our purpose?

Thanks a lot.

Read More
Answer

Thank you for contacting us.

The ab9337 recognizes proteins phosphorylated on threonine residues. It does not cross-react with phosphotyrosine.

For ab15556, this antibody recognizes serine, threonine, and tyrosine phosphorylated proteins. It may cross-react with other phosphorylated amino acids.

For ab17464, it detects phosphoserine or phosphothreonine in the context of tyrosine, tryptophan or phenylalanine at the -1 position or phenylalanine at the +1 position. The antibody does not recognize the nonphosphorylated form of these motifs, nor does it recognize other phosphoserine or phosphothreonine containing proteins and peptides.

So I probably wouldn't recommend ab17464 for your purposes, and it seems that ab15556 may be the best fit.

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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Question

we would appreciate if we should have the opportunity to test instead the ab17464 antibody, the ab15556 antibody.
Many thanks and best regards

Read More
Answer

Thank you very much for your email.
The antibody ab17464 has been qc tested for its specificity and we have found no problem with it. It is highly likely that the expressed protein you are using is not compatible with the antibodies, therefore I would suggest using the whole cell extract of cells line e.g. THP-1, calyculin A treated A431 cell lysates or mouse lung tissue extract as positive control.
We are sending you ab15556 as a free of charge replacement. The order number is 1114355. This antibody is also fully tested and guaranteed as anti Phosphoserine/threonine/tyrosine.
Please note if this antibody fails again with suggested proper positive controls then only this product is eligible for refund or replacement; we would not be able to replace it, if the product fails with recombinant protein again.
Many thanks for your cooperation in this case. Please do not hesitate to contact us, if you need further assistance.

Read More

Question

referring to our recent inquiry for the antibody anti phospho
serine/threonine (cat n° 17464), the new lot of antibody was tested.
Unfortunately, neither the new lot worked, the results was no band
detected, not even in the positive control.
The experimental condition was the same described for the previous inquiry.
What are your suggestions?
Thanks in advance
Regards

Read More
Answer

Thank you for contacting us. I am sorry to hear this antibody is not providing satisfactory results.
The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful.
Having reviewed the protocol details, I believe this product should have given satisfactory results. I apologize for the inconvenience and am pleased to offer you a refund in compensation.
Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

Read More

Question

Abcam customer care service is really helpful and cooperative. We really
appreciate and give thanks for your help.
I prefer a replacement with a different lot. I will inform you as soon as
the new lot is checked.
Regards

Read More
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 1088417.
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.

Read More

Question

as suggested from you in the previous email, a new test of the antibody
(code ab17464) was checked and the phosphorylation of the recombinant
protein was assessed by performing incubation with CHIP.

Answering to your questions, the protein that is being used is AurKA,
commercially available and purified from SF9. The protein is supposed to
be post-translationally modified as a consequence of SF9 cells production.

As you can see from the attached file, the protein is endogenously highly
phosphorylated (revealed by using anti phospho-Thr antibody from Cell
Signaling company) and dephosphorylated in the presence of 40U CHIP.

The protein was incubated in the same kinase buffer used in our previous
experiments and any step was altered.


Since this antibody was used by us in the past and it worked properly, we
can suppose that a mistake in the shipment occured.

Waiting for your istructions

Read More
Answer

Thank you for your cooperation in this case.
It may well be that this antibody in vial might have degraded or might have lost the specificity due to unknown reasons. In order to avoid any further delays in your project I am happy to offer you either free of charge replacement with a different lot or a totally different antibody.
Could you let me know how you would like to proceed?

Read More

Question

in attaching you can find the filled questionnaire, and the immuno blotting film.
Waiting for instructions from you
My best regards

Read More
Answer

Thank you for your email. I am sorry to hear that you are experiencing problems with this antibody.
I have checked the protocol which seems fine to me. However I have doubt about the recombinant proteins you have been using. Could you please give more details about the recombinant protein.
- Have you checked the phosphorylation by any other method?
- What kind of recombinant protein is being used? e.g. human RAS, RAC etc.
- Is it expressed or synthetic?
I also have further suggestion which may improve results.
- I can suggest running whole cell/ tissue lysates to check if the no band is because of antibody or recombinant protein itself?
- Running 10ug will give more prominent results.
I hope these suggestions will help to improve the results. Should the results not improves please do not hesitate to contact me.

Read More

Question

I am writing you in reference to our recent order (n. 247 / 2012), of the anti-Phosphoserine / Threonine antibody (cat n. 17464).
Yesterday (May 3th), we tried the antibody.
Unfortunately, after tree tests, we didn't obtain any signal.
For this reason, we would kindly ask for a replacement of the shipment.
Waiting for instructions from you
My best regards.

Read More
Answer

Thank you for contacting us.
I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.
I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.
I look forward to receiving your reply.

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Immunocytochemistry/ Immunofluorescence abreview for Anti-Phosphoserine/threonine antibody

 Good
Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human osteosarcoma SaOS-2)
Specification
human osteosarcoma SaOS-2
Fixative
Paraformaldehyde
Permeabilization
Yes - PBS TRITON .25%
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Read More

Abcam user community

Verified customer

Submitted May 02 2012

1-10 of 19 Abreviews or Q&A

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