Overview

  • Product nameAnti-Phospholamban antibody [2D12]
    See all Phospholamban primary antibodies
  • Description
    Mouse monoclonal [2D12] to Phospholamban
  • Tested applicationsSuitable 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

  • EpitopeThis 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

  • FunctionReversibly 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 specificityHeart muscle (at protein level).
  • Involvement in diseaseCardiomyopathy, dilated 1P
    Cardiomyopathy, familial hypertrophic 18
  • Sequence similaritiesBelongs 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 localizationEndoplasmic 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) »
  • Tang Y  et al. Nitric oxide enhances extracellular ATP induced Ca²? oscillation in HeLa cells. Arch Biochem Biophys 565:68-75 (2015). Read more (PubMed: 25461674) »

See all 17 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.

http://www....

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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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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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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"