Anti-Phospholamban antibody (ab85146)

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Overview

  • Product name

    Anti-Phospholamban antibody
    See all Phospholamban primary antibodies

  • Description

    Rabbit polyclonal to Phospholamban

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Rabbit, Horse, Cow, Dog, Pig, Macaque monkey

  • Immunogen

    Synthetic peptide corresponding to Human Phospholamban aa 1 to the C-terminus conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab124138)

  • Positive control

    • This antibody gave a positive signal in the following tissue lysates: Human Heart; Mouse Heart; Rat Heart. IHC-P: human duodenum FFPE tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab85146 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 Use a concentration of 1 µg/ml. Detects a band of approximately 6 kDa (predicted molecular weight: 6 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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

    see all

  • Alternative names

    • Cardiac phospholamban antibody
    • CMD1P antibody
    • CMH18 antibody
    • PLB antibody
    • Pln antibody
    • PPLA_HUMAN antibody
    see all

Images

  • Western blot - Anti-Phospholamban antibody (ab85146)
    Western blot - Anti-Phospholamban antibody (ab85146)
    All lanes : Anti-Phospholamban antibody (ab85146) at 1 µg/ml

    Lane 1 : Human heart tissue lysate - total protein (ab29431)
    Lane 2 : Heart (Mouse) Tissue Lysate
    Lane 3 : Heart (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 6 kDa
    Observed band size: 6 kDa


    Exposure time: 8 minutes
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phospholamban antibody (ab85146)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Phospholamban antibody (ab85146)
    IHC image of Phospholamban staining in human duodenum formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab85146, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References

This product has been referenced in:

  • Jozwiak M  et al. Concomitant systolic and diastolic alterations during chronic hypertension in pig. J Mol Cell Cardiol 131:155-163 (2019). Read more (PubMed: 31051181) »
  • Pires AL  et al. Reverse myocardial effects of intermedin in pressure-overloaded hearts: role of endothelial nitric oxide synthase activity. J Physiol 591:677-87 (2013). Read more (PubMed: 23165766) »
See all 2 Publications for this product

Publishing research using ab85146? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-2 of 2 Q&A

Question

Hi:
I am sorry to bother you.
I want to do western blot to detect the expression of phospholamban and phospho-phospholamban on guinea pig bladder smoothe muscle. I want to use the antibodies as following: ab85146(phospholamban) and ab15000(phospho-phospholamban)
Meanwhile, I have some questions and need your help.
1. I found two antibodies(ab85146 & ab86930) for phospholamban and one antibody(ab15000) for phospho-phospholamban. But I didn't find they can react with guinea pig in cross reactivity of their datasheet.
So, I want know whether the three antibodies can react with guinea pig of not.
2. I want to know the difference betweeb ab85146 and ab86930 and which one is more suitable for guinea pig.

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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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Question

Hi: I want to detect the protein expression of Phospholamban and Phospholamban(phospho S16) on Guinea pig bladder smoothe muscle, in the presence or absence of drugs by western blot.So I want to know as following about ab86930, ab85146 and ab15000 as following: 1. What is the difference between ab86930 and ab85146? Which is more suitable for guinea pig? 2. Are there other antibodies more suitable for Phospholamban and Phospholamban(phospho S16) on guinea pig? 3.Which lysate solution is best for Phospholamban and Phospholamban(phospho S16)? 4.Which protease and phosphatase inhibitors are best for the two protein? 5.Would you like give the protocal for the phosphorylation protein,such as phospho S16?

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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.

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