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Validated using a knockout cell line

Anti-ABCA1 antibody [HJ1] - BSA and Azide free (ab264550)

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Western blot - Anti-ABCA1 antibody [HJ1] - BSA and Azide free (ab264550)
  • Flow Cytometry - Anti-ABCA1 antibody [HJ1] - BSA and Azide free (ab264550)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABCA1 antibody [HJ1] - BSA and Azide free (ab264550)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABCA1 antibody [HJ1] - BSA and Azide free (ab264550)

Key features and details

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

You may also be interested in

Protein
Recombinant Human ABCA1 protein (ab125995)
Secondary
Product image
Goat Anti-Mouse IgG H&L (HRP) (ab205719)

View more associated products

Overview

  • Product name

    Anti-ABCA1 antibody [HJ1] - BSA and Azide free
    See all ABCA1 primary antibodies
  • Description

    Mouse monoclonal [HJ1] to ABCA1 - BSA and Azide free
  • Host species

    Mouse
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    WB
    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
  • General notes

    ab264550 is the carrier-free version of ab66217.

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

     

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    HJ1
  • Isotype

    IgG2b
  • Light chain type

    kappa
  • Research areas

    • Neuroscience
    • Cell Type Marker
    • Glia marker
    • Astrocyte marker
    • Cardiovascular
    • Lipids / Lipoproteins
    • Lipid Metabolism
    • Cholesterol Metabolism
    • Signal Transduction
    • Metabolism
    • Lipid metabolism
    • Cardiovascular
    • Atherosclerosis
    • Lipoprotein metabolism
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of lipids and lipoproteins
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Lipid metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Cholesterol Metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Lipid and lipoprotein metabolism
    • Lipoprotein metabolism
    • Metabolism
    • Types of disease
    • Heart disease

Associated products

  • Alternative Versions

    • Anti-ABCA1 antibody [HJ1] (ab66217)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772)
    • Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
  • Recombinant Protein

    • Recombinant Human ABCA1 protein (ab125995)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab264550 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
IHC-P
Human
WB
Human
All applications
Rat
Application Abreviews Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 254 kDa (predicted molecular weight: 254 kDa).

Abcam recommends using 3% Milk as the blocking agent.

IHC-P
Use a concentration of 0.2 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt
Use 2µg for 106 cells.

(methanol fixed cells)

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 254 kDa (predicted molecular weight: 254 kDa).

Abcam recommends using 3% Milk as the blocking agent.

IHC-P
Use a concentration of 0.2 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt
Use 2µg for 106 cells.

(methanol fixed cells)

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

Target

  • Function

    cAMP-dependent and sulfonylurea-sensitive anion transporter. Key gatekeeper influencing intracellular cholesterol transport.
  • Tissue specificity

    Widely expressed, but most abundant in macrophages.
  • Involvement in disease

    Defects in ABCA1 are a cause of high density lipoprotein deficiency type 1 (HDLD1) [MIM:205400]; also known as analphalipoproteinemia or Tangier disease (TGD). HDLD1 is a recessive disorder characterized by absence of high density lipoprotein (HDL) cholesterol from plasma, accumulation of cholesteryl esters, premature coronary artery disease (CAD), hepatosplenomegaly, recurrent peripheral neuropathy and progressive muscle wasting and weakness.
    Defects in ABCA1 are a cause of high density lipoprotein deficiency type 2 (HDLD2) [MIM:604091]; also known as familial hypoalphalipoproteinemia (FHA). HDLD2 is inherited as autosomal dominant trait. It is characterized by moderately low HDL cholesterol, predilection toward premature coronary artery disease (CAD) and a reduction in cellular cholesterol efflux.
  • Sequence similarities

    Belongs to the ABC transporter superfamily. ABCA family.
    Contains 2 ABC transporter domains.
  • Domain

    Multifunctional polypeptide with two homologous halves, each containing an hydrophobic membrane-anchoring domain and an ATP binding cassette (ABC) domain.
  • Post-translational
    modifications

    Phosphorylation on Ser-2054 regulates phospholipid efflux.
    Palmitoylation by DHHC8 is essential for membrane localization.
  • Cellular localization

    Membrane.
  • Target information above from: UniProt accession O95477 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 19 Human
    • Entrez Gene: 11303 Mouse
    • Entrez Gene: 313210 Rat
    • Omim: 600046 Human
    • SwissProt: O95477 Human
    • SwissProt: P41233 Mouse
    • Unigene: 429294 Human
    • Unigene: 277376 Mouse
    • Unigene: 148916 Rat
    see all
  • Alternative names

    • ABC 1 antibody
    • ABC Transporter 1 antibody
    • ABC-1 antibody
    • ABC1 antibody
    • ABCA 1 antibody
    • ABCA1 antibody
    • ABCA1_HUMAN antibody
    • ATP binding Cassette 1 antibody
    • ATP binding cassette sub family A ABC1 member 1 antibody
    • ATP binding cassette sub family A member 1 antibody
    • ATP binding Cassette Transporter 1 antibody
    • ATP-binding cassette 1 antibody
    • ATP-binding cassette sub-family A member 1 antibody
    • ATP-binding cassette transporter 1 antibody
    • CERP antibody
    • Cholesterol efflux regulatory protein antibody
    • FLJ14958 antibody
    • HDLDT1 antibody
    • Membrane bound antibody
    • MGC164864 antibody
    • MGC165011 antibody
    • TD antibody
    • TGD antibody
    see all

Images

  • Western blot - Anti-ABCA1 antibody [HJ1] - BSA and Azide free (ab264550)
    Western blot - Anti-ABCA1 antibody [HJ1] - BSA and Azide free (ab264550)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab264550).

  • Flow Cytometry - Anti-ABCA1 antibody [HJ1] - BSA and Azide free (ab264550)
    Flow Cytometry - Anti-ABCA1 antibody [HJ1] - BSA and Azide free (ab264550)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab264550).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABCA1 antibody [HJ1] - BSA and Azide free (ab264550)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABCA1 antibody [HJ1] - BSA and Azide free (ab264550)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab264550).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABCA1 antibody [HJ1] - BSA and Azide free (ab264550)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABCA1 antibody [HJ1] - BSA and Azide free (ab264550)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab264550).

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (0)

Publishing research using ab264550? Please let us know so that we can cite the reference in this datasheet.

ab264550 has not yet been referenced specifically in any publications.

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