Anti-ABCA1 antibody [HJ1] (ab66217)
Key features and details
- Mouse monoclonal [HJ1] to ABCA1
- Suitable for: WB, 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
Suitable for: WB, Flow Cyt, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
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
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 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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Some batches contain 6.97% L-Arginine as a stabilizing agent. For lot-specific buffer information, please contact our Scientific Support team. -
Concentration information loading...
-
Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
HJ1 -
Isotype
IgG2b -
Light chain type
kappa -
Research areas
- 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
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab66217 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 |
---|---|---|
WB | (1) |
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. |
Flow Cyt | (1) |
Use 2µg for 106 cells.
(methanol fixed cells) ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody. |
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.
|
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. |
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. |
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. |
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
modificationsPhosphorylation on Ser-2054 regulates phospholipid efflux.
Palmitoylation by DHHC8 is essential for membrane localization. -
Cellular localization
Membrane. - 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
-
Alternative names
- ABC 1 antibody
- ABC Transporter 1 antibody
- ABC-1 antibody
see all
Images
-
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)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)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. -
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. -
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.
Datasheets and documents
-
SDS download
-
Datasheet download
References (15)
ab66217 has been referenced in 15 publications.
- Wang X et al. Cholesterol and saturated fatty acids synergistically promote the malignant progression of prostate cancer. Neoplasia 24:86-97 (2022). PubMed: 34954451
- Lu Q et al. Selective activation of ABCA1/ApoA1 signaling in the V1 by magnetoelectric stimulation ameliorates depression via regulation of synaptic plasticity. iScience 25:104201 (2022). PubMed: 35479414
- Hu J et al. A reduction of Syndecan-4 in macrophages promotes atherosclerosis by aggravating the proinflammatory capacity of macrophages. J Transl Med 20:319 (2022). PubMed: 35842658
- Wang Y et al. 27-Hydroxycholesterol-Induced Dysregulation of Cholesterol Metabolism Impairs Learning and Memory Ability in ApoE ε4 Transgenic Mice. Int J Mol Sci 23:N/A (2022). PubMed: 36232940
- Zhu L et al. Expressing the Human Cholesteryl Ester Transfer Protein Minigene Improves Diet-Induced Fatty Liver and Insulin Resistance in Female Mice. Front Physiol 12:799096 (2021). PubMed: 35082691