Recombinant
RabMAb

Recombinant Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)

Reviews (3)Q&A (4)References (15)

Overview

  • Product name

    Anti-Apolipoprotein A I antibody [EP1368Y]
    See all Apolipoprotein A I primary antibodies

  • Description

    Rabbit monoclonal [EP1368Y] to Apolipoprotein A I

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Sandwich ELISA, IHC-FoFr, WB, IP, Flow Cyt, IHC-P, ICC/IF, ELISAmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human Apolipoprotein A I aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • WB: Human liver and fetal lysates IHC-P: Human liver tissue ICC/IF: HepG2 cells FC: HepG2 cells IP: HepG2 cell lysates, Human fetal liver tissue lysate

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Applications

Our Abpromise guarantee covers the use of ab52945 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
Sandwich ELISA Use a concentration of 0 - 0.1 mg/ml. Can be paired for Sandwich ELISA with Goat polyclonal to Apolipoprotein A I (HRP) (ab20784).
IHC-FoFr Use at an assay dependent concentration.
WB 1/1000. Detects a band of approximately 31 kDa (predicted molecular weight: 31 kDa).

For unpurified use at 1/20000 dilution.
IP 1/60.
Flow Cyt 1/20.
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
ICC/IF 1/100 - 1/250.
ELISA Use at an assay dependent concentration.

Antibody conc. = 0 - 1000 ng/ml

Target

  • Function

    Participates in the reverse transport of cholesterol from tissues to the liver for excretion by promoting cholesterol efflux from tissues and by acting as a cofactor for the lecithin cholesterol acyltransferase (LCAT). As part of the SPAP complex, activates spermatozoa motility.

  • Tissue specificity

    Major protein of plasma HDL, also found in chylomicrons. Synthesized in the liver and small intestine.

  • Involvement in disease

    Defects in APOA1 are a cause of high density lipoprotein deficiency type 2 (HDLD2) [MIM:604091]; also known as familial hypoalphalipoproteinemia (FHA). Inheritance is autosomal dominant.
    Defects in APOA1 are a cause of the low HDL levels observed in high density lipoprotein deficiency type 1 (HDLD1) [MIM:205400]; also known as analphalipoproteinemia or Tangier disease (TGD). HDLD1 is a recessive disorder characterized by the absence of plasma HDL, accumulation of cholesteryl esters, premature coronary artery disease, hepatosplenomegaly, recurrent peripheral neuropathy and progressive muscle wasting and weakness. In HDLD1 patients, ApoA-I fails to associate with HDL probably because of the faulty conversion of pro-ApoA-I molecules into mature chains, either due to a defect in the converting enzyme activity or a specific structural defect in Tangier ApoA-I.
    Defects in APOA1 are the cause of amyloid polyneuropathy-nephropathy Iowa type (AMYLIOWA) [MIM:107680]; also known as amyloidosis van Allen type or familial amyloid polyneuropathy type III. AMYLIOWA is a hereditary generalized amyloidosis due to deposition of amyloid mainly constituted by apolipoprotein A1. The clinical picture is dominated by neuropathy in the early stages of the disease and nephropathy late in the course. Death is due in most cases to renal amyloidosis. Severe peptic ulcer disease can occurr in some and hearing loss is frequent. Cataracts is present in several, but vitreous opacities are not observed.
    Defects in APOA1 are a cause of amyloidosis type 8 (AMYL8) [MIM:105200]; also known as systemic non-neuropathic amyloidosis or Ostertag-type amyloidosis. AMYL8 is a hereditary generalized amyloidosis due to deposition of apolipoprotein A1, fibrinogen and lysozyme amyloids. Viscera are particularly affected. There is no involvement of the nervous system. Clinical features include renal amyloidosis resulting in nephrotic syndrome, arterial hypertension, hepatosplenomegaly, cholestasis, petechial skin rash.

  • Sequence similarities

    Belongs to the apolipoprotein A1/A4/E family.

  • Post-translational
    modifications

    Palmitoylated.
    Phosphorylation sites are present in the extracelllular medium.

  • Cellular localization

    Secreted.
  • Information by UniProt

  • Database links

    • Alternative names

      • Apo-AI antibody
      • ApoA I antibody
      • ApoA-I antibody
      • APOA1 antibody
      • APOA1_HUMAN antibody
      • Apolipoprotein A-I(1-242) antibody
      • Apolipoprotein A1 antibody
      • Apolipoprotein AI antibody
      • Apolipoprotein of high density lipoprotein antibody
      • ApolipoproteinAI antibody
      • Brp14 antibody
      • high density lipoprotein uptake antibody
      • Ltw1 antibody
      • Lvtw1 antibody
      • MGC117399 antibody
      • Sep1 antibody
      • Sep2 antibody
      see all

    Images

    • Western blot - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)
      Western blot - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)
      Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945) at 1/1000 dilution (Purified) + HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 15 µg

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

      Predicted band size: 31 kDa
      Observed band size: 31 kDa

    • Western blot - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)
      Western blot - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)
      Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945) at 1/1000 dilution (Purified) + Human liver lysates at 15 µg

      Secondary
      Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

      Predicted band size: 31 kDa
      Observed band size: 31 kDa

    • Western blot - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)
      Western blot - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)
      Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945) at 1/20000 dilution ((unpurified)) + fetal liver lysate at 10 µg

      Secondary
      goat anti-rabbit HRP at 1/2000 dilution

      Predicted band size: 31 kDa
      Observed band size: 31 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver tissue sections labeling Apolipoprotein A I with purified ab52945 at 1/100 dilution (1.95 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Immunocytochemistry/ Immunofluorescence - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)
      Immunocytochemistry/ Immunofluorescence - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)

      Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Apolipoprotein A I with purified ab52945 at 1/250 dilution (0.8 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 µg/ml) dilution. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Flow Cytometry - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)
      Flow Cytometry - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)

      Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Apolipoprotein A I with purified ab52945 at 1/20 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    • Immunoprecipitation - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)
      Immunoprecipitation - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)

      ab52945 (purified) at 1/20 dilution (1ug) immunoprecipitating Apolipoprotein A I in Human fetal liver lysates.
      Lane 1: Human fetal liver lysates 10ug
      Lane 2 (+): ab52945 & Human fetal liver lysates
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52945 in Human fetal liver lysates
      For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    • ELISA - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)
      ELISA - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)

      Direct ELISA antigen dose-response curve using purified ab52945 at 0-1000ng/mL. Antigen (apo AI peptide) concentration of 1000ng/mL. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)

      ab52945 (unpurified) at 1/100 dilution staining Apolipoprotein A I in human liver by Immunohistochemistry, Paraffin embedded tissue.

    • Immunocytochemistry/ Immunofluorescence - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)
      Immunocytochemistry/ Immunofluorescence - Anti-Apolipoprotein A I antibody [EP1368Y] (ab52945)

      ab52945 (unpurified) at 1/100 dilution staining Apolipoprotein A I in HEPG2 cells by Immunofluorescence.

    References

    This product has been referenced in:

    • Camacho J  et al. Brain ApoA-I, ApoJ and ApoE Immunodetection in Cerebral Amyloid Angiopathy. Front Neurol 10:187 (2019). Read more (PubMed: 30918495) »
    • Mazzeo A  et al. Functional analysis of miR-21-3p, miR-30b-5p and miR-150-5p shuttled by extracellular vesicles from diabetic subjects reveals their association with diabetic retinopathy. Exp Eye Res 184:56-63 (2019). Read more (PubMed: 31002820) »
    See all 15 Publications for this product

    Publishing research using ab52945? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review

    Filter by Application

    Filter by Species

    Filter by Ratings

    1-3 of 3 Abreviews

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Apolipoprotein A I antibody [EP1368Y]

     Inconclusive
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Small Intestine)
    Antigen retrieval step
    Heat mediated
    Permeabilization
    No
    Specification
    Small Intestine
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More

    Arnab Chattopadhyay

    Verified customer

    Submitted Oct 31 2018

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Apolipoprotein A I antibody [EP1368Y]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Mouse Tissue sections (Brain)
    Permeabilization
    Yes - 0.1 M PBS with 1% Triton X
    Specification
    Brain
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
    Fixative
    Paraformaldehyde
    Read More

    Dr. Ruma Raha-Chowdhury

    Verified customer

    Submitted Feb 05 2016

    Western blot abreview for Anti-Apolipoprotein A I antibody [EP1368Y]

     Average
    Abreviews
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Brain lysate)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    30 µg
    Specification
    Brain lysate
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Dr. Ruma Raha-Chowdhury

    Verified customer

    Submitted Jan 28 2016

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com