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RecombinantRabMAb

Recombinant Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
  • Immunocytochemistry/ Immunofluorescence - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
  • Western blot - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
  • Flow Cytometry (Intracellular) - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
  • Immunoprecipitation - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
  • Western blot - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
  • Western blot - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
  • Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR24118-2] to Angiotensinogen - BSA and Azide free
  • Suitable for: WB, IHC-P, IP, ICC/IF, Flow Cyt (Intra)
  • Reacts with: Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free
    See all Angiotensinogen primary antibodies

  • Description

    Rabbit monoclonal [EPR24118-2] to Angiotensinogen - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, IP, ICC/IF, Flow Cyt (Intra)more details
    Unsuitable for: IHC-Fr

  • Species reactivity

    Reacts with: Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human plasma, lung, liver and kidney tissue lysates; Rat plasma; Rat lung, kidney and liver tissue tissue lysate; HepG2 and Huh7 whole cell lysates. Flow Cyt (intra): HepG2 cells. ICC/IF: HepG2 cells. IHC-P: Rat kidney tissue; Human placenta, liver, and kidney tissue. IP: HepG2 whole cell lysate.

  • General notes

    ab276137 is the carrier-free version of ab276132.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab276137 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 at an assay dependent concentration. Detects a band of approximately 48-70 kDa (predicted molecular weight: 53 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
Notes
WB
Use at an assay dependent concentration. Detects a band of approximately 48-70 kDa (predicted molecular weight: 53 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
Application notes
Is unsuitable for IHC-Fr.

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)

      This data was developed using ab276132, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Angiotensinogen with ab276132 at 1/500 (0.854 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on rat kidney (PMID: 28716988). The section was incubated with ab276132 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.

    • Immunocytochemistry/ Immunofluorescence - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
      Immunocytochemistry/ Immunofluorescence - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)

      This data was developed using ab276132, the same antibody clone in a different buffer formulation.

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 100% Methanol permeabilized HepG2 cells labelling Angiotensinogen with ab276132 at 1/100 (4.27 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000  dilution (Green). Confocal image showing cytoplasmic staining in HepG2 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

    • Western blot - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
      Western blot - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
      All lanes : Anti-Angiotensinogen antibody [EPR24118-2] (ab276132) at 1/1000 dilution

      Lane 1 : HepG2 (human hepatocellar carcinoma epithelial cell), whole cell lysate
      Lane 2 : Huh7 (human hepatocellar carcinoma epithelial cell), whole cell lysate
      Lane 3 : Human lung tissue lysate
      Lane 4 : Human liver tissue lysate
      Lane 5 : Human kidney tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 53 kDa
      Observed band size: 48-70 kDa why is the actual band size different from the predicted?



      This data was developed using ab276132, the same antibody clone in a different buffer formulation. 

      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      Angiotensinogen undergoes glycosylation. The molecular weight observed is consistent with what has been described in literature(PMID: 9694881).

      Lanes 1 & 2 of the blot were developed using a higher sensitivity ECL substrate.

      Exposure times: Lane 1, 2: 59 seconds; Lane 3: 3.25 seconds;Lane 4, 5: 15 seconds

    • Flow Cytometry (Intracellular) - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
      Flow Cytometry (Intracellular) - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)

      This data was developed using ab276132, the same antibody clone in a different buffer formulation. 

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling Angiotensinogen with ab276132 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). 

      A Goat anti rabbit IgG (Alexa Fluor®488, ab150077) at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Immunoprecipitation - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
      Immunoprecipitation - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)

      This data was developed using ab276132, the same antibody clone in a different buffer formulation.

      Angiotensinogen was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate with ab276132 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab276132 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate 10 ug

      Lane 2: ab276132 IP in HepG2 whole cell lysate

      Lane 3:  Rabbit monoclonal IgG (ab172730) instead of ab276132 in HepG2 whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 23 seconds.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)

      This data was developed using ab276132, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling Angiotensinogen with ab276132 at 1/500 (0.854 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human placenta (PMID: 21215447). The section was incubated with ab276132 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.

    • Western blot - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
      Western blot - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
      All lanes : Anti-Angiotensinogen antibody [EPR24118-2] (ab276132) at 1/1000 dilution

      Lane 1 : Rat lung tissue lysate
      Lane 2 : Rat liver tissue lysate
      Lane 3 : Rat kidney tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

      Predicted band size: 53 kDa
      Observed band size: 48-70 kDa why is the actual band size different from the predicted?



      This data was developed using ab276132, the same antibody clone in a different buffer formulation. 

      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      Angiotensinogen undergoes glycosylation. The molecular weight observed is consistent with what have been described in literature(PMID: 9694881).

      This blot was developed using a higher sensitivity ECL substrate.

      Exposure time: 127 seconds.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)

      This data was developed using ab276132, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Angiotensinogen with ab276132 at 1/500 (0.854 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human kidney. The section was incubated with ab276132 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.

    • Western blot - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
      Western blot - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
      All lanes : Anti-Angiotensinogen antibody [EPR24118-2] (ab276132) at 1/1000 dilution

      Lane 1 : Human plasma
      Lane 2 : Rat plasma

      Lysates/proteins at 20 µl per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/100000 dilution

      Predicted band size: 53 kDa
      Observed band size: 48-70 kDa why is the actual band size different from the predicted?



      This data was developed using ab276132, the same antibody clone in a different buffer formulation. 

      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

      Angiotensinogen undergoes glycosylation. The molecular weight observed is consistent with what have been described in literature(PMID: 9694881).

      Exposure times: 

      Lane 1: 15 seconds
      Lane 2: 3.25 seconds

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)

      This data was developed using ab276132, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Angiotensinogen with ab276132 at 1/500 (0.854 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human liver. The section was incubated with ab276132 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.

    • Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)
      Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free (ab276137)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab276137? Please let us know so that we can cite the reference in this datasheet.

    ab276137 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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    Flow Cytometry abreview for Anti-Angiotensinogen antibody [EPR24118-2] - BSA and Azide free

     Average
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (Immortalized Human Astrocytes)
    Permeabilization
    Yes - CyTOF permeabilization buffer
    Specification
    Immortalized Human Astrocytes
    Preparation
    Cell harvesting/tissue preparation method: Trypsin-EDTA for 2-3 minutes
    Sample buffer: PriGrow 4
    Fixation
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Sep 22 2021

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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