Validated using a knockout cell line

Anti-ABCA1 antibody [AB.H10] (ab18180)

Knockout Tested Mouse monoclonal ABCA1 antibody [AB.H10]. Validated in WB, ELISA, IHC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Chicken, Cow, Human. Cited in 81 publication(s).

Reviews (17)Q&A (36)References (88)

Overview

  • Product name
    Anti-ABCA1 antibody [AB.H10]
    See all ABCA1 primary antibodies
  • Description
    Mouse monoclonal [AB.H10] to ABCA1
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-P, ICC/IF, Flow Cyt, WB, ELISAmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Cow, Human
  • Immunogen

    Recombinant fragment corresponding to Human ABCA1 aa 1800-2260.
    Database link: O95477

  • Positive control
    • testis, liver, and brain tissue (negative control: muscle tissue)
  • General notes

    This product was changed from ascites to tissue culture supernatant on 5th February 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Applications

Our Abpromise guarantee covers the use of ab18180 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
IHC-P 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use 1-2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB 1/200 - 1/500. Predicted molecular weight: 254 kDa.

Abcam recommends using BSA as the blocking agent and 80-100 ug of lysate.
ELISA Use at an assay dependent concentration.

Direct ELISA, where a 96-well plate was coated with recombinant ABCA1 C-terminal protein. The antibody was then added to the wells and a goat anti-mouse IgG HRP conjugate was used for colour development. For ABCA1 quantification, a pair of anti-ABCA1 antibodies should be used to formulate a Sandwich ELISA and with a presence of an ABCA1 protein Standard. Optimal dilution to be determined by the end user.

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.
  • Information by UniProt
  • Database links
    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 [AB.H10] (ab18180)
    Western blot - Anti-ABCA1 antibody [AB.H10] (ab18180)
    Western blot analysis of ABCA1 induction by cholesterol (+) in human fibroblast cells from different patients (1,2,3,4). Simultaneous bloting with anti-actin antibody was used for protein loading control. Western blot analysis of ABCA1 induction by cholesterol (+) in human fibroblast cells from different patients (1,2,3,4). Simultaneous blotting with anti-actin antibody was used for protein loading control.
  • Flow Cytometry - Anti-ABCA1 antibody [AB.H10] (ab18180)
    Flow Cytometry - Anti-ABCA1 antibody [AB.H10] (ab18180)
    Overlay histogram showing HepG2 cells stained with ab18180 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab18180, 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 IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Western blot - Anti-ABCA1 antibody [AB.H10] (ab18180)
    Western blot - Anti-ABCA1 antibody [AB.H10] (ab18180)
    All lanes : Anti-ABCA1 antibody [AB.H10] (ab18180) at 1/1000 dilution

    Lanes 1 & 3 & 5 : Wild type mouse liver tissue lysate
    Lanes 2 & 4 & 6 : Knockout mouse liver tissue lysate

    Lysates/proteins at 100 µg per lane.

    Predicted band size: 254 kDa
    Observed band size: 254 kDa



    The lower bands are from the secondary anti-mouse antibody reacting to the endogenous mouse antibodies from the tissue.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABCA1 antibody [AB.H10] (ab18180)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ABCA1 antibody [AB.H10] (ab18180)
    Immunoreactivity of pancreas tissue for ABCA1 protein. Left panel: strong immunopositivity of exocrine glandular cells of the pancreas, especially in the basal region (arrows). Interstitial microvascular cells are apparently negative. Cells within the islet were weakly positive (data not shown). Right panel: the negative control. Hematoxylin counterstain.

References

This product has been referenced in:
  • Huang Z  et al. G protein pathway suppressor 2 (GPS2) links inflammation and cholesterol efflux by controlling lipopolysaccharide-induced ATP-binding cassette transporter A1 expression in macrophages. FASEB J 33:1631-1643 (2019). Read more (PubMed: 30153049) »
  • Fan J  et al. CKIP-1 limits foam cell formation and inhibits atherosclerosis by promoting degradation of Oct-1 by REG?. Nat Commun 10:425 (2019). Read more (PubMed: 30683852) »
See all 88 Publications for this product

Publishing research using ab18180? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 53 Abreviews or Q&A

Western blot abreview for Anti-ABCA1 antibody [AB.H10]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Sheep Cell lysate - whole cell (Ovine Thecal Cells)
Gel Running Conditions
Reduced Denaturing (10% PAG)
Loading amount
10 µg
Specification
Ovine Thecal Cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Read More

Miss. Barbara Makela

Verified customer

Submitted Jun 22 2017

Western blot abreview for Anti-ABCA1 antibody [AB.H10]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (liver)
Gel Running Conditions
Reduced Denaturing (7.5)
Loading amount
25 µg
Specification
liver
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Nov 24 2016

Immunocytochemistry/ Immunofluorescence abreview for Anti-ABCA1 antibody [AB.H10]

 Average
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Primary astrocyte culture)
Permeabilization
Yes - 0.1M PBS with 1% Triton X
Specification
Primary astrocyte culture
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Methanol
Read More

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Sep 06 2016

Western blot abreview for Anti-ABCA1 antibody [AB.H10]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Rabbit Tissue lysate - whole (Synovial membrane)
Gel Running Conditions
Reduced Denaturing (8 %)
Loading amount
35 µg
Specification
Synovial membrane
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Jun 13 2016

Flow Cytometry abreview for Anti-ABCA1 antibody [AB.H10]

 Good
Abreviews
Application
Flow Cytometry
Fixation
Formaldehyde
Permeabilization
Yes - BD PERMEABILISATION SOLUTION
Sample
Mouse Cell (liver cells)
Specification
liver cells
Gating Strategy
CD24 POSITIVE
Preparation
Cell harvesting/tissue preparation method: single cell suspension
Sample buffer: 0.1%PBS-BSA
Read More

Abcam user community

Verified customer

Submitted Dec 10 2013

Flow Cytometry abreview for Anti-ABCA1 antibody [AB.H10]

 Good
Abreviews
Application
Flow Cytometry
Fixation
Formaldehyde
Permeabilization
Yes - BD Permeabilisation Solution
Sample
Rat Cell (Liver cells)
Specification
Liver cells
Gating Strategy
CD24 Positive
Preparation
Cell harvesting/tissue preparation method: Single cell suspension
Sample buffer: 0.1% PBS-BSA
Read More

Abcam user community

Verified customer

Submitted Dec 05 2013

Flow Cytometry abreview for Anti-ABCA1 antibody [AB.H10]

 Good
Abreviews
Application
Flow Cytometry
Fixation
Formaldehyde
Permeabilization
Yes - BD PERMEABILISATION SOLUTION
Sample
Human Cell (HEP G2)
Specification
HEP G2
Gating Strategy
CD24 POSITIVE
Preparation
Cell harvesting/tissue preparation method: single cell suspension
Sample buffer: 0.1%PBS-BSA
Read More

Mr. brahmi manali

Verified customer

Submitted Dec 04 2013

Western blot abreview for Anti-ABCA1 antibody [AB.H10]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Pig Tissue lysate - whole (small intestine)
Loading amount
30 µg
Specification
small intestine
Gel Running Conditions
Reduced Denaturing (7)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 6% · Temperature: 20°C
Read More

Mr. Dongil Kim

Verified customer

Submitted Apr 23 2013

Question

Thank you for your kind replacement of the antibody! I was able to test it on human prostate cancer cell line PC3 lysed with RIPA buffer (last lane of the samples indicated by arrow) and my other prostate cancer cell lines prepared before. The membrane was first detected with your new antibody and then reprobed with FASN. As you can see from the images, your new antibody can only show a low molecular weight background band. FASN antibody clearly detects the target protein with several non-specific bands. The pictures in two different pages represents different exposure time. Therefore, I am afraid that your new antibody does not work either. If you can provide a positive control, I am willing to repeat experiment again. If your positive control works, it means that your antibodies work. I am happy to keep them and improve my sample preparation.

Read More
Answer


Yes, I can send you as a one time exception a positive control lysate (rat liver (ab29413), https://www.abcam.com/liver-rat-whole-cell-lysate-normal-tissue-ab29413.html). This has been tested with ab66217 and gave strong bands.

Also I would suggest to check the gel after transfer as some of the high MW proteins might have not completely transfered onto the membrane. You can check this by staining the gel with Coomassie Blue. if you see bands in the gel at 200 kDa and higher, you might want to adjust your transfer conditions so that all high MW protein are being transfered.

Also, avoid boiling of your samples before loading them onto the gel. Warming your sample to 50-60 degree Celsius is sufficient for denaturation in loading buffer.

Again, FASN is a cytoplasmic protein and such is much easier to work with than ABCA1 which is very hydrophobic, glycosylated and of high MW.

This article and link might be of help as well: http://www.sciencedirect.com/science/article/pii/S0003269708006829
http://www.scientistsolutions.com/t14685-membrane+protein+western+blot.html

Read More

Question

I saw that you changed the pictures from WB of ABAC1 in your datasheet. As you know I didn´t find the protein at , the actual, predicted molecular weigth of 220Kda of the previous 294Kda. My question is, the lower band at +|- 60Kda ( you also found) is correlated with the upper band? The size of the bands are correlated for the same condition?

Read More
Answer

In this image the lower band is actin which has been recognized by and anti-actin antibody. This has been used as a loading control to normalize the levels of protein detected by confirming that protein loading is the same across the gel. The expression levels of the loading control should not vary between the different sample lanes.

Read More

1-10 of 53 Abreviews or Q&A

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