Overview

  • Product name
    Anti-Phospholamban antibody [2D12]
    See all Phospholamban primary antibodies
  • Description
    Mouse monoclonal [2D12] to Phospholamban
  • Tested applications
    Suitable for: ICC/IF, Inhibition Assay, IHC-P, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Rabbit, Chicken, Guinea pig, Dog, Human, Pig
    Predicted to work with: Cow
  • Immunogen

    Synthetic peptide corresponding to Dog Phospholamban aa 2-25.
    Sequence:

    DKVQYLTRSAIRRASTIEMPQQAR

  • Epitope
    This antibody recognizes an epitope between amino acid residues 9-17 of canine PLB.

Properties

Applications

Our Abpromise guarantee covers the use of ab2865 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 1/10 - 1/100.
Inhibition Assay Use at an assay dependent concentration.
IHC-P 1/200.
IP Use at an assay dependent concentration.
WB 1/500 - 1/5000.

This antibody detects a 25 kDa protein under non-reducing conditions representing the pentameric form of PLB and a 5 kDa protein under reducing conditions representing the monomeric form of PLB in canine cardiac extracts.

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

Anti-Phospholamban antibody [2D12] images

  • Ab2865 staining human normal liver. Staining is localized to the cytoplasm.
    Left panel: with primary antibody at 1/1000. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. 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 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, 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 we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

  • IF/ICC image of Phospholamban antibody (ab2865) staining hES cell derived cardiomyocytes (red). The cells were fixed in paraformaldehyde and blocked in 4% goat serum for 1 hour at 25°C. The Phospholamban antibody was used at a concentration of 1:250, for 16 hours at 4°C.

    See Abreview

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human skeletal muscle tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing Phospholamban ab2865 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human heart tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing Phospholamban ab2865 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

References for Anti-Phospholamban antibody [2D12] (ab2865)

This product has been referenced in:
  • Te Rijdt WP  et al. Phospholamban p.Arg14del cardiomyopathy is characterized by phospholamban aggregates, aggresomes, and autophagic degradation. Histopathology 69:542-50 (2016). Read more (PubMed: 26970417) »
  • Ahmad F  et al. Regulation of sarcoplasmic reticulum Ca2+ ATPase 2 (SERCA2) activity by phosphodiesterase 3A (PDE3A) in human myocardium: phosphorylation-dependent interaction of PDE3A1 with SERCA2. J Biol Chem 290:6763-76 (2015). Read more (PubMed: 25593322) »

See all 19 Publications for this product

Product Wall

Thank you for your phone call this afternoon.

As discussed, here is the mous-on-mouse protocol I mentioned. This is primarily concerned with background generated when using a mouse antibody when staining tissue from mouse.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (foetal cardiac myocytes)
Specification
foetal cardiac myocytes
Fixative
Methanol
Permeabilization
Yes - Triton 0.1%
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
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Verified customer

Submitted Apr 27 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (hESC-derived cardiomyocyte)
Specification
hESC-derived cardiomyocyte
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100 in PBS
Blocking step
Goat serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
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Submitted Nov 15 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - other (cardiac tissue)
Loading amount
10 µg
Specification
cardiac tissue
Gel Running Conditions
Reduced Denaturing (4-12% nupage bis-tris)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Submitted Sep 17 2010

Application
Western blot
Sample
African Green Monkey Tissue lysate - whole (hamster heart tissue)
Loading amount
25 µg
Specification
hamster heart tissue
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
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Submitted Mar 17 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (C2C12 cells)
Loading amount
20 µg
Specification
C2C12 cells
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
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Submitted Jun 05 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (Rat heart tissue)
Loading amount
25 µg
Specification
Rat heart tissue
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
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Submitted Jun 05 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rabbit Tissue lysate - whole (rabbit heart tissue)
Loading amount
20 µg
Specification
rabbit heart tissue
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
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Submitted Jun 05 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (ventricular tissue lysates)
Loading amount
20 µg
Specification
ventricular tissue lysates
Gel Running Conditions
Non-reduced Denaturing (4~12% gradient NuPAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
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Submitted Jul 05 2008

Thank you for your enquiry. The immunogen for this anti-phospholamban antibody corresponds to amino acid residues 2-25 of canine PLB. The amino acids in question are included in this sequence, however, they were unphosphorylated when used as the immun...

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