Overview

  • Product nameAnti-SERCA2 ATPase antibody [IID8]
    See all SERCA2 ATPase primary antibodies
  • Description
    Mouse monoclonal [IID8] to SERCA2 ATPase
  • SpecificityDetects Sarcoplasmic or Endoplasmic Reticulum Calcium 2 (SERCA2) ATPase. This antibody does not discriminate between the two isoforms. By Western blot, this antibody detects an ~110 kDa protein representing SERCA2 ATPase from canine skeletal muscle triad preparations. Immunofluorescence staining of SECRA2 ATPase in rabbit skeletal muscle results in strong labeling of the entire type I (slow) myofiber consistent with sarcoplasmic reticulum localization. This antibody is not recommended for Western blot detection of rat SERCA2.
  • Tested applicationsSuitable for: ICC/IF, IHC-Fr, WB, ICC, Inhibition Assay, ELISA, IP, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Rat, Sheep, Rabbit, Guinea pig, Hamster, Cow, Dog, Human, Pig
    Predicted to work with: Non Human Primates, Amphibians
  • Immunogen

    Full length native protein (purified) corresponding to Dog SERCA2 ATPase. Purified canine cardiac sarcoplasmic reticulum.

  • Positive control
    • canine skeletal muscle triad preparations

Properties

Applications

Our Abpromise guarantee covers the use of ab2817 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 Use at an assay dependent concentration.
IHC-Fr 1/500.
WB 1/2500. Detects a band of approximately 100 kDa.
ICC Use at an assay dependent concentration.
Inhibition Assay Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 1 µg/ml.
Flow Cyt Use 1µg for 106 cells.

Target

  • FunctionThis magnesium-dependent enzyme catalyzes the hydrolysis of ATP coupled with the translocation of calcium from the cytosol to the sarcoplasmic reticulum lumen. Isoform 2 is involved in the regulation of the contraction/relaxation cycle.
  • Tissue specificityIsoform 1 is widely expressed in smooth muscle and nonmuscle tissues such as in adult skin epidermis, with highest expression in liver, pancreas and lung, and intermediate expression in brain, kidney and placenta. Also expressed at lower levels in heart and skeletal muscle. Isoforms 2 and 3 are highly expressed in the heart and slow twitch skeletal muscle. Expression of isoform 3 is predominantly restricted to cardiomyocytes and in close proximity to the sarcolemma. Both isoforms are mildly expressed in lung, kidney, liver, pancreas and placenta. Expression of isoform 3 is amplified during monocytic differentiation and also observed in the fetal heart.
  • Involvement in diseaseDefects in ATP2A2 are a cause of acrokeratosis verruciformis (AKV) [MIM:101900]; also known as Hopf disease. AKV is a localized disorder of keratinization, which is inherited as an autosomal dominant trait. Its onset is early in life with multiple flat-topped, flesh-colored papules on the hands and feet, punctate keratoses on the palms and soles, with varying degrees of nail involvement. The histopathology shows a distinctive pattern of epidermal features with hyperkeratosis, hypergranulosis, and acanthosis together with papillomatosis. These changes are frequently associated with circumscribed elevations of the epidermis that are said to resemble church spires. There are no features of dyskeratosis or acantholysis, the typical findings in lesions of Darier disease.
    Defects in ATP2A2 are the cause of Darier disease (DD) [MIM:124200]; also known as Darier-White disease (DAR). DD is an autosomal dominantly inherited skin disorder characterized by loss of adhesion between epidermal cells (acantholysis) and abnormal keratinization. Patients with mild disease may have no more than a few scattered keratotic papules or subtle nail changes, whereas those with severe disease are handicapped by widespread malodorous keratotic plaques. In a few families, neuropsychiatric abnormalities such as mild mental retardation, schizophrenia, bipolar disorder and epilepsy have been reported. Stress, UV exposure, heat, sweat, friction, and oral contraception exacerbate disease symptoms. Prevalence has been estimated at 1 in 50000. Clinical variants of DD include hypertrophic, vesicobullous, hypopigmented, cornifying, zosteriform or linear, acute and comedonal subtypes. Comedonal Darier disease (CDD) is characterized by the coexistence of acne-like comedonal lesions with typical Darier hyperkeratotic papules on light-exposed areas. At histopathologic level, CDD differs from classic DD in the prominent follicular involvement and the presence of greatly elongated dermal villi.
  • Sequence similaritiesBelongs to the cation transport ATPase (P-type) (TC 3.A.3) family. Type IIA subfamily.
  • Post-translational
    modifications
    Nitrated under oxidative stress. Nitration on the two tyrosine residues inhibits catalytic activity.
  • Cellular localizationEndoplasmic reticulum membrane. Sarcoplasmic reticulum membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • AT2A2_HUMAN antibody
    • Atp2a2 antibody
    • ATP2B antibody
    • ATPase Ca++ transporting cardiac muscle slow twitch 2 antibody
    • Calcium pump 2 antibody
    • Calcium-transporting ATPase sarcoplasmic reticulum type antibody
    • Calcium-transporting ATPase sarcoplasmic reticulum type slow twitch skeletal muscle isoform antibody
    • Cardiac Ca2+ ATPase antibody
    • DAR antibody
    • DD antibody
    • Endoplasmic reticulum class 1/2 Ca(2+) ATPase antibody
    • MGC45367 antibody
    • Sarcoplasmic/endoplasmic reticulum calcium ATPase 2 antibody
    • SERCA 2 antibody
    • SERCA2 antibody
    • serca2a antibody
    • slow twitch skeletal muscle isoform antibody
    • SR Ca(2+)-ATPase 2 antibody
    see all

Anti-SERCA2 ATPase antibody [IID8] images

  • ab28217 (2µg/ml) staining calpain S1 in renal medulla
    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 buffers EDTA pH 9.0 . 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 we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
  • ab2817 (1µg/ml) staining SERCA2 ATPase in human heart (left ventricle), using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining of cardiomyocytes.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 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 ab2817 stained MCF7 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 (ab2817, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse 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.

  • All lanes : Anti-SERCA2 ATPase antibody [IID8] (ab2817) at 1 µg/ml

    Lane 1 : Human skeletal muscle tissue lysate - total protein (ab29330)
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 99 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 30 kDa,44 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 8 minutes
  • Overlay histogram showing HepG2 cells stained with ab2817 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2817, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human heart tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) 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 SERCA2 ATPase (ab2817) 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 liver tissue. 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 SERCA2 ATPase (ab2817) 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 tonsil tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) 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 SERCA2 ATPase (ab2817) 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.
  • Immunofluorescent analysis of SERCA2 ATPase using SERCA2 ATPase Monoclonal antibody (IID8) ab2817 shows staining in U251 glioma cells. SERCA2 ATPase staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing SERCA2 ATPase ab2817 at a dilution of 1:100-1:200 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunofluorescent analysis of SERCA2 ATPase using SERCA2 ATPase Monoclonal antibody (IID8) ab2817 shows staining in A549 cells. SERCA2 ATPase staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing SERCA2 ATPase ab2817 at a dilution of 1:100-1:200 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunofluorescent analysis of SERCA2 ATPase using SERCA2 ATPase Monoclonal antibody (IID8) ab2817 shows staining in HeLa cells. SERCA2 ATPase staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing SERCA2 ATPase ab2817 at a dilution of 1:100-1:200 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

References for Anti-SERCA2 ATPase antibody [IID8] (ab2817)

This product has been referenced in:
  • Wright MF  et al. Mechanisms of intracellular calcium homeostasis in developing and mature bovine corpora lutea. Biol Reprod 90:55 (2014). WB, IHC-P ; Cow . Read more (PubMed: 24501170) »
  • Karolczak J  et al. Myosin VI in skeletal muscle: its localization in the sarcoplasmic reticulum, neuromuscular junction and muscle nuclei. Histochem Cell Biol 139:873-85 (2013). Read more (PubMed: 23275125) »

See all 19 Publications for this product

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Selon nos informations vous avez connu quelques difficultés avec ab2817 et avez contacté notre service scientifique.

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Je suis désolée d'apprendre que ab2817 ne vous donne pas de bons résultats en Western blot sur du tissu humain ou murin.

Veuillez trouver ci-joint un questionnaire discuté au téléphone qui nous permet de regrouper le plus d'informations poss...

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Thank you for contacting us. I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly. I am attaching ou...

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Thank you for your reply. From the information you have provided, the alignment with the Danio sounds high, and although we cannot guarantee it without laboratory testing, we would suggest the antibody should detect in this fish. I can ...

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Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. Thank you for taking the time to complete our technical questionnaire. Our mouse monoclonal [IID8] to SERCA2 ATPase (ab2817) has been applied ...

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Thank you for your enquiry. Regarding ab2817 and ab2861: These monoclonal antibodies have not been epitope mapped, therefore the epitope could be anywhere along the full length of the protein. I did an alignment of Human SERCA2 with the Xenopus ...

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I'm sorry to hear you are having problems with ab2817 and ab6495. Your detailed protocol was very useful to understand the source of the problem and it is clear your protocol and modifications were very thorough. My suspicions therefore lied with y...

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