Overview

  • Product name
    Anti-Phospholamban (phospho S16) antibody
    See all Phospholamban primary antibodies
  • Description
    Rabbit polyclonal to Phospholamban (phospho S16)
  • Host species
    Rabbit
  • Specificity
    Multiple bands can be observed in addition to the 25kD pentamer for PLB. Monomeric, dimeric, trimeric forms, etc. can be observed if the preparations are boiled before SDS-PAGE. This antibody also can display non-specific bands at 75 and 100kD in some lysate preparations. Note: Do not boil cardiac preparation. Boiling may reduce the 25kD pentamer in to tri-, di- or monomers. In addition, phosphorylation may disrupt the pentamer into dimers and monomers.
  • Tested applications
    Suitable for: IHC-P, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Hamster, Cow, Human, Pig, Chinese hamster
    Predicted to work with: Dog
  • Immunogen

    KLH-conjugated, synthetic peptide corresponding to amino acids 14-25 of human cardiac phospholamban (

    RA[pS]TIEMPQQAR-C

    ) with an added C-terminal cysteine. The immunizing sequence is identical in mouse, rat, rabbit, pig and dog.

  • Positive control
    • WB: Bovine cardiac preparation treated with PKA..

Properties

Applications

Our Abpromise guarantee covers the use of ab15000 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
IHC-P Use at an assay dependent concentration.
WB Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 6.24 kDa.
IHC-Fr Use at an assay dependent concentration.

Target

  • Function
    Reversibly inhibits the activity of ATP2A2 in cardiac sarcoplasmic reticulum by decreasing the apparent affinity of the ATPase for Ca(2+). Modulates the contractility of the heart muscle in response to physiological stimuli via its effects on ATP2A2. Modulates calcium re-uptake during muscle relaxation and plays an important role in calcium homeostasis in the heart muscle. The degree of ATP2A2 inhibition depends on the oligomeric state of PLN. ATP2A2 inhibition is alleviated by PLN phosphorylation.
  • Tissue specificity
    Heart muscle (at protein level).
  • Involvement in disease
    Cardiomyopathy, dilated 1P
    Cardiomyopathy, familial hypertrophic 18
  • Sequence similarities
    Belongs to the phospholamban family.
  • Post-translational
    modifications
    Phosphorylation by PKA abolishes the inhibition of ATP2A2-mediated calcium uptake. Phosphorylated at Thr-17 by CaMK2, and in response to beta-adrenergic stimulation. Phosphorylation by DMPK may stimulate sarcoplasmic reticulum calcium uptake in cardiomyocytes.
  • Cellular localization
    Endoplasmic reticulum membrane. Sarcoplasmic reticulum membrane. Mitochondrion membrane. Membrane. Colocalizes with HAX1 at the endoplasmic reticulum (PubMed:17241641). Colocalizes with DMPK a the sarcoplasmic reticulum (PubMed:15598648).
  • Information by UniProt
  • Database links
  • Alternative names
    • Cardiac phospholamban antibody
    • CMD1P antibody
    • CMH18 antibody
    • PLB antibody
    • Pln antibody
    • PPLA_HUMAN antibody
    see all

Images

  • All lanes : Anti-Phospholamban (phospho S16) antibody (ab15000) at 0.5 µg/ml

    Lane 1 : Bovine cardiac preparation treated with PKA
    Lane 2 : Untreated bovine cardiac preparation

    Predicted band size: 6.24 kDa
    Additional bands at: 14 kDa (possible multimer), 23 kDa (possible multimer)

  • ab15000 staining Phospholamban (phospho S16) in Rat heart tissue by Immunohistochemistry (Frozen sections). The sections were fixed in Acetone prior to blocking with neat SuperBlock Blocking Buffer for 20 minutes at 23°C. The primary antibody was diluted 1/50 in PBS and incubated with the sample for 12 hours at 4°C. ab6720 - Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Biotin) was used as the secondary antibody, diluted 1/100.

    See Abreview

  • Formalin-fixed, paraffin-embedded pig heart tissue stained for Phospholamban (phospho S16) using ab15000 at 1/150 dilution in immunohistochemical analysis. 

    See Abreview

References

This product has been referenced in:
  • Wu Y  et al. Cardiac Protection of Valsartan on Juvenile Rats with Heart Failure by Inhibiting Activity of CaMKII via Attenuating Phosphorylation. Biomed Res Int 2017:4150158 (2017). WB . Read more (PubMed: 28536695) »
  • Silveira AC  et al. Exercise Training Restores Cardiac MicroRNA-1 and MicroRNA-29c to Nonpathological Levels in Obese Rats. Oxid Med Cell Longev 2017:1549014 (2017). WB . Read more (PubMed: 29138674) »
See all 18 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Sample
Mouse Cell (myocyte)
Specification
myocyte
Permeabilization
No
Fixative
Acetone

Abcam user community

Verified customer

Submitted Jun 17 2013

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (15)
Sample
Human Tissue lysate - whole (Heart)
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 10 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Pig Tissue lysate - whole (Heart)
Loading amount
7.5 µg
Specification
Heart
Gel Running Conditions
Reduced Denaturing (14% PAGE)
Blocking step
membrane blocking solution (Invitrogen) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Dec 19 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (heart)
Specification
heart
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: universal decloaker
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Nov 27 2012

Answer

Thank you for contacting us. We have not tested these antibodies in guinea pig and therefore cannot guarantee they will work, however there is high homology between the immunogen and the guinea pig sequence for all three. Normally we predictlikely reactivity if there is 85% homology of the immunogen with the species, and for ab15000 there is 91%, ab85146 is 90% and ab86930 is 93%. The two phospholambin antibodies ab85146 and ab86930 are very similar. They are both rabbit polyclonal purified antibodies and have similar likelihood of reactiving with guinea pig. They do have difference concentrations and volumes (ab85146 is provided at 1ug/ml and 100 ul; ab86930 is 0.5 ug/ml and 200 ul) and have different immunogens. Either would be a good choice.


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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Read More

Answer

Thank you for contacting Abcam.

I would be very happy to help answer your questions:

1 - What is the difference between ab86930 and ab85146? Which is more suitable for guinea pig?

- Both antibodies target different areas of the Phospholamban protein. For a lot of targets, Abcam offers multiply antibodies, that either target different domains, or are more suited to different applications, depending on the customers’ needs.

I cannot find the guinea pig sequence for Phospholamban, therefore I cannot make a recommendation as to whether either of these antibodies would work for that species. If you could please supply the guinea pig sequence, I would be happy to do the alignment and see if there is a chance that either will work for you.

2 - Are there other antibodies more suitable for Phospholamban and Phospholamban(phospho S16) on guinea pig?

- Again, without the guinea pig sequence, I cannot confidently answer this question.

3 - Which lysate solution is best for Phospholamban and Phospholamban(phospho S16)?

- From the information available on the webpages, it looks as though mouse, rat, human and bovine cardiac tissue either treated with or without PKA would be suitable lysates.

4 - Which protease and phosphatase inhibitors are best for the two protein?

- This would be determined on your exact experiment at hand, below is a link to some protease inhibitors, however if you require phospahatse inhibitors then, we unfortunately do not carry those in our catalogue:

https://www.abcam.com/Protease-Inhibitor-Cocktail-ab65621.html



5. Would you like give the protocol for the phosphorylation protein,such as phospho S16?

- For ab15000 was treated with PKA to induce phosphorylation.

Please let me know if there is anything else I can help you with, or if you would like to me to perform the alignment between the guinea pig sequence and the immunogen sequences for these antibodies.

Read More

Answer

I am sorry to hear that you have been experiencing problems using this product in the application that you wish. Unfortunately, the attachment has not come through successfully. Could you please resend it to me? It is much appreciated.

In order to assess the quality of our products I would ask that you complete a brief questionnaire relating to the application used. Often it is possible to make suggestions that may help resolve problems experienced using a particular product.

As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

All our customer feedback, including complaints are monitored weekly by our in house technical support team. If a product is at fault the technical support team will consider removing the product from our catalogue in order to avoid future customer inconvenience.

Please could you provide some further details of the protocol used and complete the following form (attached as a word document).

Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (heart)
Specification
heart
Fixative
Acetone
Permeabilization
No
Blocking step
SuperBlock Blocking Buffer as blocking agent for 20 minute(s) · Concentration: 100% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jan 19 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (heart tissue)
Loading amount
20 µg
Specification
heart tissue
Gel Running Conditions
Reduced Denaturing (15%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Nov 04 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Chinese Hamster Cell lysate - whole cell (CHO cell line)
Loading amount
20 µg
Specification
CHO cell line
Gel Running Conditions
Reduced Denaturing (15% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 17 2009

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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