Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1374Y] to Apolipoprotein E - Low endotoxin, Azide free
  • Suitable for: ICC, WB, IHC-P, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free
    See all Apolipoprotein E primary antibodies

  • Description

    Rabbit monoclonal [EP1374Y] to Apolipoprotein E - Low endotoxin, Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC, WB, IHC-P, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK293T, HepG2 and HeLa cell lysates; Human liver cancer tissue lysate. Flow Cyt: HepG2 cells. IP: HepG2 whole cell lysate and Human serum.

  • General notes

    ab184845 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    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.

    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.

Images

  • Immunocytochemistry - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)
    Immunocytochemistry - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)

    This data was developed using ab52607, the same antibody clone in a different buffer formulation.
    Immunocytochemistry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Apolipoprotein E with Purified ab52607 at 1:50 dilution (2.0 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) 1:200 (2.5 µg/ml). 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.

  • Western blot - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)
    Western blot - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)
    All lanes : Anti-Apolipoprotein E antibody [EP1374Y] (ab52607) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : APOE knockout HEK293T cell lysate
    Lane 3 : HepG2 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 36 kDa
    Observed band size: 36 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab52607).

    Lanes 1-3: Merged signal (red and green). Green - ab52607 observed at 36 kDa. Red - loading control ab7291 observed at 50 kDa. 

     ab52607 Anti-Apolipoprotein E antibody [EP1374Y] was shown to specifically react with Apolipoprotein E in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266546 (knockout cell lysate ab256838) was used.  Wild-type and Apolipoprotein E knockout samples were subjected to SDS-PAGE.  ab52607 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively.  Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)
    Western blot - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)
    All lanes : Anti-Apolipoprotein E antibody [EP1374Y] (ab52607) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : TGFA knockout HeLa cell lysate
    Lane 3 : Human liver cancer tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 36 kDa
    Observed band size: 36 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab52607).

    Lanes 1-3: Merged signal (red and green). Green - ab52607 observed at 36 kDa. Red - loading control ab7291 observed at 50 kDa.

    ab52607 Anti-Apolipoprotein E antibody [EP1374Y] was shown to specifically react with Apolipoprotein E in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265166 (knockout cell lysate ab257739) was used. Wild-type and Apolipoprotein E knockout samples were subjected to SDS-PAGE. ab52607 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

     

  • Flow Cytometry - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)
    Flow Cytometry - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)

    Flow cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Apolipoprotein E (red) with purified ab52607 at a 1/250 dilution (10ug/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti rabbit IgG (Alexa Fluor®488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52607).

  • Immunoprecipitation - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)
    Immunoprecipitation - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)

    This data was developed using ab52607, the same antibody clone in a different buffer formulation.
    Purified ab52607 at 1/20 dilution (0.5µg) immunoprecipitating Apolipoprotein E in HepG2 whole cell lysate.
    Lane 1 (input): HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab52607 + HepG2 whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52607 in HepG2 whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 36 kDa

  • Immunoprecipitation - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)
    Immunoprecipitation - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)

    This data was developed using ab52607, the same antibody clone in a different buffer formulation.
    Purified ab52607 at 1/20 dilution (0.5µg) immunoprecipitating Apolipoprotein E in Human serum.
    Lane 1 (input): Human serum 10µg
    Lane 2 (+): ab52607 + Human serum.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52607 in Human serum.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 36 kDa

  • Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)
    Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845)

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