Overview

  • Product name

    Anti-AACT antibody [EPR17088-68]
    See all AACT primary antibodies
  • Description

    Rabbit monoclonal [EPR17088-68] to AACT
  • Host species

    Rabbit
  • Specificity

    This antibody recognizes a different epitope of AACT compared to ab204514.
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Full length native protein (purified) corresponding to Human AACT aa 1 to the C-terminus.
    Database link: P01011

  • Positive control

    • WB: Human plasma (untreated and PNGase F treated) and blood; Human fetal spleen lysate. IHC-P: Human tonsil tissue. ICC/IF: Raji cells. IP: Human plasma lysate.
  • 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab205198 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. Detects a band of approximately 50-70 kDa (predicted molecular weight: 47 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/50.
IP 1/40.

Target

  • Function

    Although its physiological function is unclear, it can inhibit neutrophil cathepsin G and mast cell chymase, both of which can convert angiotensin-1 to the active angiotensin-2.
  • Tissue specificity

    Plasma. Synthesized in the liver. Like the related alpha-1-antitrypsin, its concentration increases in the acute phase of inflammation or infection. Found in the amyloid plaques from the hippocampus of Alzheimer disease brains.
  • Involvement in disease

    Defects in SERPINA3 may be a cause of chronic obstructive pulmonary disease (COPD) [MIM:107280].
  • Sequence similarities

    Belongs to the serpin family.
  • Domain

    The reactive center loop (RCL) extends out from the body of the protein and directs binding to the target protease. The protease cleaves the serpin at the reactive site within the RCL, establishing a covalent linkage between the carboxyl group of the serpin reactive site and the serine hydroxyl of the protease. The resulting inactive serpin-protease complex is highly stable.
  • Cellular localization

    Secreted.
  • Information by UniProt
  • Database links

  • Alternative names

    • SERPINA3 antibody
    • AACT antibody
    • AACT_HUMAN antibody
    • ACT antibody
    • alpha 1 Antichymotrypsin antibody
    • Alpha-1-antichymotrypsin His-Pro-less antibody
    • Antichymotrypsin antibody
    • Cell growth inhibiting gene 24/25 protein antibody
    • Cell growth-inhibiting gene 24/25 protein antibody
    • GIG24 antibody
    • GIG25 antibody
    • Growth inhibiting protein 24 antibody
    • Growth inhibiting protein 25 antibody
    • MGC88254 antibody
    • Serine (or cysteine) proteinase inhibitor clade A (alpha 1 antiproteinase, antitrypsin) member 3 antibody
    • Serine (or cysteine) proteinase inhibitor clade A member 3 antibody
    • Serine proteinase inhibitor clade A member 3 antibody
    • Serpin A3 antibody
    • Serpin family A member 3 antibody
    • Serpin peptidase inhibitor clade A (alpha 1 antiproteinase antitrypsin) member 3 antibody
    • SERPINA3 antibody
    see all

Images

  • Anti-AACT antibody [EPR17088-68] (ab205198) at 1/2000 dilution + Human plasma at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 47 kDa
    Observed band size: 50-70 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The expression profile observed is consistent with what has been described in the literature (PMID: 9880565).

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling AACT with ab205198 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab205198) at 1/500 dilution. Cytoplasmic staining on human tonsil tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji (Human Burkitt's lymphoma cell line) cells labeling AACT with ab205198 at 1/50 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and weak nuclear staining on Raji cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab205198 at 1/50 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Anti-AACT antibody [EPR17088-68] (ab205198) at 1/1000 dilution + His-Tagged Human SERPINA3 (aa 24 to 324) recombinant protein 100ng

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 47 kDa
    Observed band size: 33 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Diluting buffer and concentration 5% NFDM/TBST

  • Anti-AACT antibody [EPR17088-68] (ab205198) at 1/1000 dilution + Human blood at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 47 kDa
    Observed band size: 50-70 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

     

  • Anti-AACT antibody [EPR17088-68] (ab205198) at 1/1000 dilution + Human fetal spleen lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 47 kDa
    Observed band size: 50-70 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-AACT antibody [EPR17088-68] (ab205198) at 1/2000 dilution

    Lane 1 : Untreated Human plasma
    Lane 2 : Human plasma treated with PNGase F

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/100000 dilution

    Predicted band size: 47 kDa
    Observed band size: 50-70 kDa why is the actual band size different from the predicted?


    Exposure time: 4 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    De-glycosylation treatment with PNGase, demonstrates a molecular weight shift from 50-70kDa to 46kDa. This result is consistent with what has been described in literature the literature (PMID:) 9880565.

  • AACT was immunoprecipitated from 1 mg of Human plasma lysate with ab205198 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab205198 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.

    Lane 1: Human plasma lysate, 10 μg (Input).

    Lane 2: ab205198 IP in Human plasma lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab205198 in Human plasma lysate.

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

    Exposure time: 30 seconds.

     

References

ab205198 has not yet been referenced specifically in any publications.

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