Anti-PDCD4 (phospho S457) antibody (ab74141)

Thank you for your comments.

The paper deals with localization of PDCD4 according to its phosphorylation state.

PDCD4 can be phosphorylated on Ser67, via mTor, or phosphorylated on Ser457 via PI3K.

Only phosphorylation on Ser457 leads to its localization in the cytoplasm. Under starvation condition ser457 is phosphorylated, while mTor phosphorylation of Ser67 happens with serum addition.

In the abreview, the problem was a very weak staining. I used a confocal microscope with high laser power in order to get staining signal.

I attach a picture with staiing I did with the ProSci antibody (not for the phosphorylated form). I think the difference is clear.

ANSWER:

Thanks for your reply.

If you don't feel the vial of ab74141 you received is working as expected, I'd be happy to offer a replacement, credit or refund if you have submitted this review within about 6 months of receiving the antibody.

In your reply, please include the PO number or Abcam order reference number associated with the purchase of ab74141 so I can process your request.

AbID: 74141, Anti-PDCD4 (phospho S457) antibody Rating: Poor Image: Anti-PDCD4 (phospho S457) antibody for Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) (Mouse) Sample: Species: Mouse Type: Tissue sections Specification: embryonic pancreas Application: Application: Fixative: Paraformaldehyde Antigen retrieval step: Heat mediated - Buffer/Enzyme Used: glycine buffer pH 9 Permeabilization: No Blocking step: cas-block (invitrogen) as blocking agent for 15 mins at 20°C Other detail: Dilution: 1/100 Incubation time: 18 hours at 4°C Diluent: cas-block Secondary Antibody: Name: Non-Abcam Antibody was used: jackson 111-165-144 Conjugation: Cy3 ® Dilution: 1/300 Additional Data: Additional Notes: abreview code 42639 The staining was very weak. The antigen retrieval caused too much background. Using the same staining procedure W/O antigen retrieval gave a very weak and granular result. (file attached) the same slides, stained with ProSci PDCD4 a/b gave much better staining

ANSWER:

Thank you for submitting your Abreview. I hope you have been notified it has been published on our website. I wanted to follow up with you regarding your review as your rating of 'Poor' indicated you were unhappy with the performance of ab74141. Please note that, although ab74141 is predicted to react with mouse PDCD4 based on the similariity of the immunogen sequence to mouse protein, it has not yet been tested for mouse reactivity. However, in investigating this issue I found a reference which I believe shows similar punctate, nuclear staining pattern in cultured cells. Please see Figure 3 in the following reference for this data: http://cancerres.aacrjournals.org/content/65/24/11282.long I look forward to hearing from you after you've had a chance to look at the data.

The customer's details are:  

Customer name: xxxxx Customer email address:xxxxxx Customers institution/company: xxxxx

Thanks in advance for sending the discount code.  

ANSWER:

Thank you for providing those extra details. The discount code generated is : xxxxx, expiration date: xxxxxxx

I am very pleased to hear your customer would like to accept our offer and test ab74141 in mouse IHC. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for mouse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site:

www.abcam.com/Abreviews

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways:

1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. The terms and conditions applicable to this offer can be found here:

www.abcam.com/collaborationdiscount

Dear Tech Support Team,  

Our customer is willing to test ab74141 in IHC-P with mouse sections. Are you interested in this type of the information for the Abreview promotion (The customer buys certain antibodies and upload Abreview on Abcam's website and then we give 100% credit note for this antibody to buy next antibody)?                               

Thanks in advance for your assistance and reply.                       

Have a nice week,    

ANSWER:

Thank you for your interest in the testing discount.

We would be interested in your customer testing ab74141 in IHC-P with mouse sections. If you could please provide the following details I will generate a discount code for your customer to use:

Customer name: Customer email address: Customers institution/company:

Many thanks for your assistance.