Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ABCA1 antibody [HJ1] (ab66217)

Key features and details

  • Mouse monoclonal [HJ1] to ABCA1
  • Suitable for: WB, ICC, Flow Cyt, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG2b

Overview

  • Product name

    Anti-ABCA1 antibody [HJ1]
    See all ABCA1 primary antibodies

  • Description

    Mouse monoclonal [HJ1] to ABCA1

  • Host species

    Mouse

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC
    Mouse
    IHC-P
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment corresponding to ABCA1 (N terminal). 50 kDa N-terminal extracellular loop of ABCA1
    Database link: O95477

  • Positive control

    • IHC-P: Liver parenchyma and liver tissue. Flow: HepG2 cells. WB: Hap1 whole cell lysate. ICC; Mouse primary neurone cells

  • General notes

    This monoclonal antibody to ABCA1 has been knockout validated in Western blot. The expected band for ABCA1 was observed in wild type cells and the band was not seen in knockout cells, although other non-specific bands were also seen. The data are shown on this datasheet.

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Immunocytochemistry - Anti-ABCA1 antibody [HJ1] (ab66217)
    Immunocytochemistry - Anti-ABCA1 antibody [HJ1] (ab66217)

    Immunocytochemistry analysis of mouse primary neurone cells labelling MGLUR2 with ab66217 at 1/100 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150113) at 1/1000 was used as the secondary antibody (green). Cells were counterstained with Anti-MAP2 rabbit monoclonal antibody (ab11267) at 1/500 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) at 1/1000 dilution (red). Nuclear DNA was labelled with DAPI (blue). 

    Confocal image showing cytoplasmic staining in mouse primary neuron cells.

    Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.

  • Western blot - Anti-ABCA1 antibody [HJ1] (ab66217)
    Western blot - Anti-ABCA1 antibody [HJ1] (ab66217)
    All lanes : Anti-ABCA1 antibody [HJ1] (ab66217) at 1 µg/ml

    Lane 1 : Brain (Mouse) Tissue Lysate
    Lane 2 : Brain (Rat) Tissue Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : Liver (Mouse) Tissue Lysate
    Lane 5 : Liver (Rat) Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 254 kDa
    Observed band size: 254 kDa
    Additional bands at: 70 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 4 minutes


    Abcam recommends using milk as the blocking agent. This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab66217 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABCA1 antibody [HJ1] (ab66217)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABCA1 antibody [HJ1] (ab66217)
    ab66217 (4µg/ml) staining ABCA1 in human liver using an automated system (DAKO Autostainer Plus). Using this protocol there is moderate cell membrane staining throughout the liver parenchyma.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABCA1 antibody [HJ1] (ab66217)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABCA1 antibody [HJ1] (ab66217)
    IHC image of ABCA1 staining in human liver formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab66217, 0.2µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • Flow Cytometry - Anti-ABCA1 antibody [HJ1] (ab66217)
    Flow Cytometry - Anti-ABCA1 antibody [HJ1] (ab66217)
    Overlay histogram showing HepG2 cells stained with ab66217 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab66217, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HepG2 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

    Please note that Abcam does not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  • Western blot - Anti-ABCA1 antibody [HJ1] (ab66217)
    Western blot - Anti-ABCA1 antibody [HJ1] (ab66217)

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: Empty
    Lane 3: ABCA1 knockout (KO) HAP1 whole cell lysate (20 µg)

     

    Lanes 1 - 3: Merged signal (red and green). Green - ab66217 observed at 240 kDa. Red - loading control, ab176560, observed at 50 kDa.

     

    ab66217 detected the expected band for ABCA1 in wild type HAP1 cells and the band was not seen in ABCA1 knockout HAP1 cells, although additional non-specific bands were also seen. Wild-type and ABCA1 knockout samples were subjected to SDS-PAGE. Ab66217 and ab176560 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1 µg/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

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

Alternative products to Anti-ABCA1 antibody [HJ1] (ab66217)

© 2021 Astana Biomed Group LLP. All rights reserved.