Overview

  • Product name
  • Description
    Rabbit polyclonal to PDE4D
  • Host species
    Rabbit
  • Specificity
    ab14613 labels all PDE4D proteins including PDE4D1, PDE4D2, PDE4D3, PDE4D4, PDE4D5 variants. Does not cross react with PDE4A, PDE4B, PDE4D6, PDE4D7, PDE4D8, PDE4D9 or PDE4C. The antibody has not been tested against endogenous PDE4D, only recombinant proteins and we therefore cannot guarantee it will detect endogenous PDE4.
  • Tested applications
    Suitable for: WB, IP, ICC, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Hamster, Human
  • Immunogen

    C-terminal cyclic peptide unique to all PDE4D subtypes variants.

  • Positive control
    • recombinant proteins (PDE4D1, PDE4D2, PDE4D3, PDE4D4, PDE4D5)

Properties

Applications

Our Abpromise guarantee covers the use of ab14613 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
WB 1/500. Can be blocked with PDE4D peptide (ab169991).
IP 1/150.
ICC 1/150.
ICC/IF Use at an assay dependent concentration. PubMed: 19759179
IHC-P Use at an assay dependent concentration.

Target

  • Function
    Hydrolyzes the second messenger cAMP, which is a key regulator of many important physiological processes.
  • Tissue specificity
    Widespread; most abundant in skeletal muscle. Isoform 6 is detected in brain. Isoform 8 is detected in brain, placenta, lung and kidney. Isoform 7 is detected in heart and skeletal muscle.
  • Pathway
    Purine metabolism; 3',5'-cyclic AMP degradation; AMP from 3',5'-cyclic AMP: step 1/1.
  • Involvement in disease
    Note=Genetic variations in PDE4D might be associated with susceptibility to stroke. PubMed:17006457 states that association with stroke has to be considered with caution.
  • Sequence similarities
    Belongs to the cyclic nucleotide phosphodiesterase family. PDE4 subfamily.
  • Post-translational
    modifications
    Isoform 3 and isoform 7 are activated by phosphorylation (in vitro), but not isoform 6. Isoform N3 and isoform 12 are phosphorylated on Ser-49, Ser-51, Ser-55 and Ser-59.
  • Cellular localization
    Cytoplasm. Membrane. Cytoplasm > cytoskeleton. Cytoplasm > cytoskeleton > centrosome. Found in the soluble fraction, associated with membranes, and associated with the cytoskeleton and the centrosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • cAMP specific 3',5' cyclic phosphodiesterase 4D antibody
    • DKFZp686M11213 antibody
    • 5''-cyclic phosphodiesterase 4D antibody
    • cAMP specific phosphodiesterase 4D antibody
    • cAMP specific phosphodiesterase PDE4D6 antibody
    • cAMP-specific 3'' antibody
    • DPDE3 antibody
    • Dunce like phosphodiesterase E3 antibody
    • FLJ97311 antibody
    • HSPDE4D antibody
    • PDE43 antibody
    • Pde4d antibody
    • PDE4D_HUMAN antibody
    • PDE4DN2 antibody
    • Phosphodiesterase 4D cAMP specific (dunce (Drosophila) homolog phosphodiesterase E3) antibody
    • Phosphodiesterase 4D cAMP specific (phosphodiesterase E3 dunce homolog Drosophila) antibody
    • Phosphodiesterase 4D cAMP specific antibody
    • Phosphodiesterase 4D, cAMP specific (dunce) antibody
    • STRK1 antibody
    see all

Images

  • ab14613 (2µg/ml) staining PDE4D in human placenta using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of the cytoplasm.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • ICC/IF image of ab14613 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab14613, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Western blot using ab14613 and positive controls for PDE4D variants.

References

This product has been referenced in:
  • Cheng H  et al. siRNA-mediated silencing of phosphodiesterase 4B expression affects the production of cytokines in endotoxin-stimulated primary cultured microglia. Exp Ther Med 12:2257-2264 (2016). WB ; Rat . Read more (PubMed: 27698721) »
  • Barbagallo F  et al. Genetically Encoded Biosensors Reveal PKA Hyperphosphorylation on the Myofilaments in Rabbit Heart Failure. Circ Res 119:931-43 (2016). Read more (PubMed: 27576469) »
See all 14 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (heart)
Permeabilization
No
Specification
heart
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Apr 27 2017

Application
Western blot
Loading amount
80 µg
Gel Running Conditions
Reduced Denaturing (4-20% gradient gel)
Sample
Mouse Cell lysate - whole cell (NIH3T3)
Specification
NIH3T3
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Xuecai Ge

Verified customer

Submitted Apr 03 2014

Answer

Thank you for your reply and for your understanding in this matter.

As requested, I am sending ab14628 as a free of charge replacement for you and you should have it on Tuesday morning. For your records the new invoice number is *.

Also, as a thank you for your patience, below is a discount code that will give 25% off 1 item on your next order with Abcam:

Discount Code: *
Exp Date: *

If there is anything else I can help you with, please let me know.

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Answer

I have tried 3 times to contact the supplier to ask them to provide those images, but they are not replying to my emails. I do apologize for the inconvenience this is causing you and I am grateful for your patience in the matter.

I believe the best way forward is to replace or refund the cost of the antibody for you. Please let me know how you would like to proceed and I will be happy to bring about a resolution to this issue.

I look forward to your reply.

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Question
Answer

Thank you for your patience in this matter.

I have heard back from the supplier in regards to ab14613 and they have tried to go back and look for old data on PDE4D antibody. They could not find the blot on HEK cells. They do have blots on other cells lines including SHS5Y neuroblastoma, rat primary neuronal cultures and in cells over expressing various PDE4D subtypes but not HEK293 cells. Those images were not included in the email they sent me.

I am very sorry about this matter and I would be happy to either replace or refund the current vial of ab14613 that you have.

Please let me know how you would like to proceed in this matter.

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Question
Answer

Thank you for contacting Abcam earlier today.

I am sorry about the western blot issues you have been having with ab14613. The antibody is covered under our Abpromise for six months and is guaranteed to work in western blot on human samples . If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

I have talked to the lab about this and they have told me one of the reason there is a band at 30kDa may be proteolytic degradation. We have checked this antibody on HEK cells and it labels 3 different PDE4D variants. Do you include protease cocktail during sample preparation? If not, I would suggest that you repeat the lysis and include the protease inhibitors.

If you already do include protease inhibitors, then I think there may be an issue with the antibody and the best way forward is to send you ab87239 as a free of charge replacement (as we discussed on the telephone).

I look forward to your reply and helping to resolve this issue.

Read More

Answer

Thank you for contacting us. I apologise for the delay, I wanted to check this case with my lab colleagues.
Regarding ab59249, the amino acid range of this immunogen is aaXXXX of human PDE4D with accession# Q08499 (http://www.uniprot.org/uniprot/Q08499).
This antibody binds to isoforms 4, 3, 5, N3, 8, 9, 7, and 12. It does not bind to isoform 10, 1, 2, and 6. According to the isoforms that this antibody binds, you would expect to see two bands, approximately 25 and 85 kDa. Because the western blot on our website is cut off at 34 kDa, we can retest this antibody to see if it does actually bind the lower isoform as well.
Although we haven’t specifically tested the antibody against all isoforms, there are other alternatives that may also recognise several forms of the protein:
For ab77080 we ran a BLAST search and found the antigen sequence in all of 9 isoforms:
https://www.abcam.com/PDE4D-antibody-ab77080.html
Ab112018 also shares 100% identity to isoforms PDE4D1-9:
https://www.abcam.com/PDE4D-antibody-ab112018.html
Ab99409 and ab87329 recognize 10 of 12 isoforms listed for SwissProt entry Q08499. The two isoforms listed for SwissProt Q08499 not recognized by either of them are sp|Q08499-7|PDE4D_HUMAN and sp|Q08499-12|PDE4D_HUMAN:
http://www.uniprot.org/uniprot/Q08499
And ab87329’s immunogen is present in 8 out of 12 of them so it should detect isoforms 1- 10.
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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Answer

I am sorry these products did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued free of charge replacements for ab14611 and ab14613with the order number 1137676.

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 contacting Abcam.

Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased:

https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount



Please contact our Scientific Support team by replying to this email prior to purchase for more information.

Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

To find out more about our Abreview system, please see the following link:

https://www.abcam.com/abreviews

I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

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Answer

Thank you for contacting us and sorry for the delay in response. I now have confirmation of the specificity of the two antibodies in question. ab14613 has been raised against a C-terminal peptide sequence of PDE4D. This peptide, contrary to what was published on the datasheet should recognise all 9 isoforms. I am sorry for this misinformation. This is as a result of science moving on and new isoforms being identified. This will now be updated on the datasheet. Similarly, ab77080 has been raised against the C-terminal amino acids (QPEACVIDDRSPDT) of human PDE4D this should also detect all 9 isoforms of PDE4D. If you have any further questions please do not hesitate to ask.

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1-10 of 15 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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