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

  • 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.
  • 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

  • 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 38 Publications for this product

Customer reviews and Q&As

1-10 of 20 Abreviews or Q&A

Application
Western blot
Sample
Mouse Tissue lysate - whole (Mouse liver)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
18 µg
Specification
Mouse liver
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Oct 15 2018

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

Abcam user community

Verified customer

Submitted Jun 26 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted May 13 2013

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.

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

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

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

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

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.

Read More

1-10 of 20 Abreviews or Q&A

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

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