Overview

  • Product name
    Anti-Angiotensinogen antibody [EPR2931]
    See all Angiotensinogen primary antibodies
  • Description
    Rabbit monoclonal [EPR2931] to Angiotensinogen
  • Host species
    Rabbit
  • Specificity
    ab108334 is raised against the Human Angiotensinogen protein. This has little homology with mouse or rat Angiotensinogen and in our hands we do not get good results when testing in these species (in western blot the band is weak or absent and at a lower molecular weight than in human samples) therefore we cannot guarantee this product for use in these species. When using this antibody for western blot we recommend blocking with 5% BSA for best results. In our hands, blocking with milk does not give optimal results in WB and some lower molecular weight binding can be observed.
  • Tested applications
    Suitable for: WB, IP, ICC/IFmore details
    Unsuitable for: Flow Cyt or IHC-P
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Angiotensinogen aa 400-500 (internal sequence). The exact sequence is proprietary.

  • Positive control
    • Human fetal artery, fetal heart, fetal liver, adult kidney, HepG2, 293T, and plasma lysates; HepG2 cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Applications

Our Abpromise guarantee covers the use of ab108334 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 1/1000 - 1/10000. Predicted molecular weight: 53 kDa.

Block with 5% BSA

IP 1/10 - 1/100.
ICC/IF 1/100 - 1/250.
  • Application notes
    Is unsuitable for Flow Cyt or IHC-P.
  • Target

    • Function
      Essential component of the renin-angiotensin system (RAS), a potent regulator of blood pressure, body fluid and electrolyte homeostasis. In response to lowered blood pressure, the enzyme renin cleaves angiotensinogen to produce angiotensin-1 (angiotensin 1-10). Angiotensin-1 is a substrate of ACE (angiotensin converting enzyme) that removes a dipeptide to yield the physiologically active peptide angiotensin-2 (angiotensin 1-8). Angiotensin-1 and angiotensin-2 can be further processed to generate angiotensin-3 (angiotensin 2-8), angiotensin-4 (angiotensin 3-8). Angiotensin 1-7 is cleaved from angiotensin-2 by ACE2 or from angiotensin-1 by MME (neprilysin). Angiotensin 1-9 is cleaved from angiotensin-1 by ACE2.
      Angiotensin-2 acts directly on vascular smooth muscle as a potent vasoconstrictor, affects cardiac contractility and heart rate through its action on the sympathetic nervous system, and alters renal sodium and water absorption through its ability to stimulate the zona glomerulosa cells of the adrenal cortex to synthesize and secrete aldosterone.
      Angiotensin-3 stimulates aldosterone release.
      Angiotensin 1-7 is a ligand for the G-protein coupled receptor MAS1 (By similarity). Has vasodilator and antidiuretic effects (By similarity). Has an antithrombotic effect that involves MAS1-mediated release of nitric oxide from platelets.
    • Tissue specificity
      Expressed by the liver and secreted in plasma.
    • Involvement in disease
      Genetic variations in AGT are a cause of susceptibility to essential hypertension (EHT) [MIM:145500]. Essential hypertension is a condition in which blood pressure is consistently higher than normal with no identifiable cause.
      Defects in AGT are a cause of renal tubular dysgenesis (RTD) [MIM:267430]. RTD is an autosomal recessive severe disorder of renal tubular development characterized by persistent fetal anuria and perinatal death, probably due to pulmonary hypoplasia from early-onset oligohydramnios (the Potter phenotype).
    • Sequence similarities
      Belongs to the serpin family.
    • Post-translational
      modifications
      Beta-decarboxylation of Asp-34 in angiotensin-2, by mononuclear leukocytes produces alanine. The resulting peptide form, angiotensin-A, has the same affinity for the AT1 receptor as angiotensin-2, but a higher affinity for the AT2 receptor.
    • Cellular localization
      Secreted.
    • Information by UniProt
    • Database links
    • Alternative names
      • Aangiotensinogen (serpin peptidase inhibitor clade A member 8) antibody
      • AGT antibody
      • AI265500 antibody
      • Alpha 1 antiproteinase antitrypsin antibody
      • Ang antibody
      • Ang I antibody
      • Ang II antibody
      • Ang III antibody
      • AngII antibody
      • Angiotensin I antibody
      • Angiotensin II antibody
      • Angiotensin III antibody
      • Angiotensin-3 antibody
      • Angiotensinogen (PAT) antibody
      • Angiotensinogen antibody
      • ANGT_HUMAN antibody
      • ANHU antibody
      • ANRT antibody
      • AT-2 antibody
      • AT-II antibody
      • Des-Asp[1]-angiotensin II antibody
      • FLJ92595 antibody
      • FLJ97926 antibody
      • MGC105326 antibody
      • PAT antibody
      • Pre angiotensinogen antibody
      • Serine (or cysteine) proteinase inhibitor antibody
      • Serpin A8 antibody
      • Serpin peptidase inhibitor clade A member 8 antibody
      • SERPINA8 antibody
      see all

    Images

    • All lanes : Anti-Angiotensinogen antibody [EPR2931] (ab108334) at 1/1000 dilution

      Lane 1 : Mouse kidney whole cell lysate
      Lane 2 : Rat kidney whole cell lysate
      Lane 3 : Human kidney whole cell lysate
      Lane 4 : Mouse liver whole cell lysate
      Lane 5 : Rat liver whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit IgG (H+L) peroxidase conjugated. at 1/2000 dilution

      Predicted band size: 53 kDa


      Exposure time: 1 minute


      Ab108334 targeting angiotensinogen in various mouse, rat and human tissues (see lane descriptions).

      Blocking buffer for the first lanes 1-5: 5% NFDM/TBST; the second lanes 1-5: BSA/TBST.

    • All lanes : Anti-Angiotensinogen antibody [EPR2931] (ab108334) at 1/1000 dilution

      All lanes : Human fetal liver whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit IgG (H+L) peroxidase conjugated. at 1/2000 dilution

      Predicted band size: 53 kDa


      Exposure time: 3 minutes


      Lane 1 blocking buffer: NFDM/TBST

      Lane 2 blocking buffer: BSA/TBST

    • All lanes : Anti-Angiotensinogen antibody [EPR2931] (ab108334) at 1/1000 dilution

      Lane 1 : Human fetal artery lysate
      Lane 2 : Human fetal heart lysate
      Lane 3 : Human fetal liver lysate
      Lane 4 : HepG2 lysate
      Lane 5 : 293T lysate
      Lane 6 : Human plasma lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 53 kDa

    • ab108334, at 1/100 dilution, staining Angiotensinogen in HepG2 cells by Immunofluorescence.

    References

    This product has been referenced in:
    • Xiang X  et al. Tuberous sclerosis complex 1-mechanistic target of rapamycin complex 1 signaling determines brown-to-white adipocyte phenotypic switch. Diabetes 64:519-28 (2015). Read more (PubMed: 25213336) »
    • Kemp JR  et al. Angiotensin II-regulated microRNA 483-3p directly targets multiple components of the renin-angiotensin system. J Mol Cell Cardiol 75:25-39 (2014). Read more (PubMed: 24976017) »
    See all 7 Publications for this product

    Customer reviews and Q&As

    1-6 of 6 Abreviews or Q&A

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunocytochemistry
    Sample
    Mouse Cultured Cells (Cardiomyocytes)
    Permeabilization
    Yes - Triton x-100, 0.03%
    Specification
    Cardiomyocytes
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Mar 13 2018

    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Cardiomyocytes)
    Gel Running Conditions
    Reduced Denaturing (10% PAGE)
    Loading amount
    25 µg
    Specification
    Cardiomyocytes
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

    Abcam user community

    Verified customer

    Submitted Sep 28 2015

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cultured Cells (human embryonic stem cells)
    Specification
    human embryonic stem cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
    Fixative
    Methanol

    Abcam user community

    Verified customer

    Submitted Jun 06 2013

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (epididymis)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate buffer
    Permeabilization
    No
    Specification
    epididymis
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    bouin's solution

    Abcam user community

    Verified customer

    Submitted Apr 09 2013

    Answer

    Thank you for contacting us. Antibodies are OK at room temperature for about 1 a week. We use this time to ship antibodies to our customers although we include icepacks in order to keep them at 4C in case of any unnecessary delays. Once received the antibody should be aliquoted and stored at -20C. At -20C the antibodies are viable for almost 10 years however it depend from product to product. Please check the datasheet for most up-to-date info regarding storage of ab108334.  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

    LOT NUMBER GR38988 ORDER NUMBER 966056 DESCRIPTION OF THE PROBLEM Non-specific band. The customer has doubt about the band size because its predicted size is 53kDa but his data showed 34kDa. SAMPLE Whole kidney lysates of mice PRIMARY ANTIBODY Concentration or dilution: 1:1000, 1:2000, 1:5000 Diluent buffer: 5% skim milk Incubation time: 2hr or overnight Incubation temperature: room temperature or 4℃ Wash Buffer: PBST (0.1% tween) Number of washes: 5min, 3 times DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED - ANTIBODY STORAGE CONDITIONS -20℃ SAMPLE PREPARATION RIPA buffer 5uM Leupeptin, 5ug/ml Aprotinin, 300 µM PMSF, 2 mM Na3VO4 5 mM DTT Boiling for ≥5 min? yes AMOUNT OF PROTEIN LOADED 30 ug/lane ELECTROPHORESIS/GEL CONDITIONS 10% SDS-PAGE gel TRANSFER AND BLOCKING CONDITIONS Type of membrane: PVDF Protein transfer verified: be checked by using ponceau solution Blocking agent and concentration: 5% skim milk Blocking time: 1, 2hr or overnight Blocking temperature: room temperature or 4℃ SECONDARY ANTIBODY Species: rabbit Isotype: IgG Reacts against: Concentration or dilution: 1:1000, 1:2000 Diluent buffer : 5% skim milk Incubation time: 1 or 1.5hr Incubation temperature: room temperature Fluorochrome or enzyme conjugate: HRP Wash Buffer: PBST (0.1% tween) Number of washes: 5min, 3 times HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? We altered several steps (blocking time, substrate concentration of ECL kit or concentration of primary or secondary antibody) ADDITIONAL NOTES I have explained the possibility of size difference but, he insist to ask his data to technical team. Check this please.

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    Answer

    Thank you for contacting Abcam Technical Team and for taking the time to provide some useful details of the experiments.   There is a very clear, single band on the customer's blot at 34 kDa, though the predicted band of precursor 53 kDa.   During maturation, Angiotensinogen is cleaved into the following 3 chains so the actual band size can be smaller than the predicted theoretical band for Angiotensinogen.   Angiotensin-1 Alternative name(s): Angiotensin I Short name=Ang I Angiotensin-2 Alternative name(s): Angiotensin II Short name=Ang II Angiotensin-3 Alternative name(s): Angiotensin III Short name=Ang III Des-Asp[1]-angiotensin II   It is also important to mention that the cleaved matured protein can be N-glycosylated which may also alter the actual size.   I hope this will be useful for your customer.

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