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
Overview
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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 HumanIHC-P HumanWB 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
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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
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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...
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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
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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Flow Cyt |
Human
|
IHC-P |
Human
|
WB |
Human
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All applications |
Rat
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Application | Abreviews | Notes |
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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 |
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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
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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
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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
see all -
Alternative names
- ABC 1 antibody
- ABC Transporter 1 antibody
- ABC-1 antibody
see all
Images
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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).
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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).
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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).
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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.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab264550 has not yet been referenced specifically in any publications.