Recombinant
RabMAb

Recombinant Anti-alpha 1 Antitrypsin antibody [EPR17087-50] (ab207303)

Overview

  • Product name

    Anti-alpha 1 Antitrypsin antibody [EPR17087-50]
    See all alpha 1 Antitrypsin primary antibodies
  • Description

    Rabbit monoclonal [EPR17087-50] to alpha 1 Antitrypsin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, IP, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Full length native protein (purified) corresponding to Human alpha 1 Antitrypsin.
    Database link: P01009

  • Positive control

    • WB: Human liver, heart, kidney, spleen and fetal liver lysates. IHC-P: Human hepatocellular carcinoma and liver tissues. ICC/IF: HepG2 and K562 cells. IP: HepG2 whole cell 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 ab207303 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
IHC-P 1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/5000. Detects a band of approximately 51, 55 kDa (predicted molecular weight: 46 kDa).
IP 1/20.
ICC/IF 1/50.

Target

  • Function

    Inhibitor of serine proteases. Its primary target is elastase, but it also has a moderate affinity for plasmin and thrombin. Irreversibly inhibits trypsin, chymotrypsin and plasminogen activator. The aberrant form inhibits insulin-induced NO synthesis in platelets, decreases coagulation time and has proteolytic activity against insulin and plasmin.
    Short peptide from AAT: reversible chymotrypsin inhibitor. It also inhibits elastase, but not trypsin. Its major physiological function is the protection of the lower respiratory tract against proteolytic destruction by human leukocyte elastase (HLE).
  • Tissue specificity

    Ubiquitous. Expressed in leukocytes and plasma.
  • Involvement in disease

    Alpha-1-antitrypsin deficiency
  • 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.
  • Post-translational
    modifications

    N-glycosylated. Differential glycosylation produces a number of isoforms. N-linked glycan at Asn-107 is alternatively di-antennary, tri-antennary or tetra-antennary. The glycan at Asn-70 is di-antennary with trace amounts of tri-antennary. Glycan at Asn-271 is exclusively di-antennary. Structure of glycans at Asn-70 and Asn-271 is Hex5HexNAc4. The structure of the antennae is Neu5Ac(alpha1-6)Gal(beta1-4)GlcNAc attached to the core structure Man(alpha1-6)[Man(alpha1-3)]Man(beta1-4)GlcNAc(beta1-4)GlcNAc. Some antennae are fucosylated, which forms a Lewis-X determinant.
    Proteolytic processing may yield the truncated form that ranges from Asp-30 to Lys-418.
  • Cellular localization

    Secreted. Endoplasmic reticulum. The S and Z allele are not secreted effectively and accumulate intracellularly in the endoplasmic reticulum and Secreted, extracellular space, extracellular matrix.
  • Information by UniProt
  • Database links

  • Alternative names

    • A1A antibody
    • A1AT antibody
    • A1AT_HUMAN antibody
    • AAT antibody
    • Alpha 1 antiproteinase antibody
    • Alpha 1 antitrypsin antibody
    • Alpha 1 antitrypsin null antibody
    • Alpha 1 protease inhibitor antibody
    • Alpha-1 protease inhibitor antibody
    • Alpha-1-antiproteinase antibody
    • alpha1 proteinase inhibitor antibody
    • Alpha1AT antibody
    • Dom1 antibody
    • PI antibody
    • PI1 antibody
    • PRO0684 antibody
    • PRO2275 antibody
    • Serine (or cysteine) proteinase inhibitor clade A member 1 antibody
    • Serine protease inhibitor 1-1 antibody
    • Serine protease inhibitor A1a antibody
    • Serpin A1 antibody
    • Serpin A1a antibody
    • Serpin peptidase inhibitor clade A member 1 antibody
    • Serpina1 antibody
    • Short peptide from AAT antibody
    • SPAAT antibody
    • Spi1-1 antibody
    see all

Images

  • All lanes : Anti-alpha 1 Antitrypsin antibody [EPR17087-50] (ab207303) at 1/5000 dilution

    Lane 1 : Human liver lysate
    Lane 2 : Human heart lysate
    Lane 3 : Human kidney lysate
    Lane 4 : Human spleen lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 46 kDa
    Observed band size: 51,55 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-alpha 1 Antitrypsin antibody [EPR17087-50] (ab207303) at 1/5000 dilution + Human fetal liver lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG. at 1/10000 dilution

    Predicted band size: 46 kDa
    Observed band size: 51,55 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling alpha 1 Antitrypsin with ab207303 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on cancer cells of human hepatocellular carcinoma 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.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue labeling alpha 1 Antitrypsin with ab207303 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human liver 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.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling alpha 1 Antitrypsin with ab207303 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HepG2 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse mAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse Alexa Fluor® 594 (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab207303 at 1/50 dilution, followed by Goat Anti-Mouse Alexa Fluor® 594 (ab150120) secondary antibody at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse mAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) seconday antibody at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling alpha 1 Antitrypsin with ab207303 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on K562 cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse mAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab207303 at 1/50 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse mAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • alpha 1 Antitrypsin was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma) whole cell lysate with ab207303 at 1/20 dilution. Western blot was performed from the immunoprecipitate using ab207303 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
    Lane 1: HepG2 whole cell lysate 10ug (Input).
    Lane 2: ab207303 IP in HepG2 whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207303 in HepG2 whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 seconds.

References

This product has been referenced in:

  • Zhao Z  et al. Silence of a1-Antitrypsin Inhibits Migration and Proliferation of Triple Negative Breast Cancer Cells. Med Sci Monit 24:6851-6860 (2018). Read more (PubMed: 30260937) »
  • Sarabhai T  et al. Serum a-1 Antitrypsin (AAT) antagonizes intrinsic apoptosis induction in neutrophils from patients with systemic inflammatory response syndrome. PLoS One 12:e0177450 (2017). Read more (PubMed: 28493974) »
See all 2 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Breast Cancer)
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: Pro-K
Permeabilization
No
Specification
Breast Cancer
Blocking step
Protein Block (ab64226) as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Feb 27 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA
Permeabilization
No
Specification
Liver
Blocking step
PB ab64226 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Feb 07 2018

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