ABCA1 Antibody

Images

 
Biological Strategies: Western Blot: ABCA1 Antibody [NB400-105] - Western Blot analysis of ABCA1 in total cell lysates of RAW264.9 cells treated with vehicle (-) or 9-cisretinoic acid and 22Rhydroxycholesterol ...read more
Biological Strategies: Western Blot: ABCA1 Antibody [NB400-105] - Detection of ABCA1 in mouse peritoneal macrophages using NB 400-105 (Lot L). ECL exposure, 1 min. Lane 4: T09 uninduced lysate Lane 5: T09 induced ...read more
Immunocytochemistry/ Immunofluorescence: ABCA1 Antibody [NB400-105] - Untreated HepG2 cells were grown to 60% confluency, and serum starved for 24 hours prior to being fixed for 10 minutes using 10% formalin and then ...read more
Immunohistochemistry-Paraffin: ABCA1 Antibody [NB400-105] - Detection of ABCA1 in human prostate epithelium showing luminal and membrane staining.
Biological Strategies: Immunocytochemistry/ Immunofluorescence: ABCA1 Antibody [NB400-105] - HepG2 cells were grown to 60% confluency, serum starved for 24 hours, and then treated with 1uM TO9 for 24 hours prior ...read more
Flow (Cell Surface): ABCA1 Antibody [NB400-105] - RAW 246.7 cells were either untreated (A) or serum starved for 24 hours, and then treated with 1uM TO9 for 24 hours (B). A surface stain was performed with ABCA1 ...read more
Western Blot: ABCA1 Antibody [NB400-105] - Analysis of ABCA1, using NB400-105. Samples: 40 ug of total cell post-nuclear lysate of raw macrophages treated with 9-cisretinoic acid and 22R-hydroxycholesterol.
Flow Cytometry: ABCA1 Antibody [NB400-105] - ABCA-1 FITC conjugated antibody of human adipose tissue macrophage subsets by flow cytometry. Image from verified customer review.
Flow Cytometry: ABCA1 Antibody [NB400-105] - ABCA1 antibody was tested at 1: 400 in HeLa cells using an Alexa Fluor 488 secondary (shown in purple). M1 is defined by unstained cells.

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Ca, Ch, Ha, MdSpecies Glossary
Applications WB, Simple Western, ChIP, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP, GS
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1 mg/ml
Validated by:
   

Biological Strategies

     

ABCA1 Antibody Summary

Immunogen
Partial peptide sequence (within residues 1100-1300) of human ABCA1 [UniProt# O95477]. Actual immunogen sequence is proprietary information.
Localization
Astrocyte Marker
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
ABCA1
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:500
  • Simple Western 1:50
  • Chromatin Immunoprecipitation
  • ELISA
  • Flow Cytometry 1:400
  • Immunocytochemistry/Immunofluorescence 1:100
  • Immunohistochemistry 1:200
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin 1:200
  • Immunoprecipitation 1:10-1:500
  • Gel Supershift Assay
Application Notes
This ABCA1 antibody has been used for Flow Cytometry and ICC (PMID: 21501868), IP (PMID: 21846716; 21106522). Use in Gel supper shift assay reported in scientific literature (PMID 15684432). In Western blot a band representing ABCA1 is observed at approx. 220 kDa. Additional non-specific bands are seen at lower molecular weights, but do not interfere with the ABCA1 signal. Chromatin Immunoprecipitation and ELISA were also reported in scientific literature.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Charge.
The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Theoretical MW
220 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Positive Control
Raw 264.7 Whole Cell - T0901317 treated Kit (NBP1-30159)
Control Peptide
ABCA1 Peptide (NB400-105PEP)
Reviewed Applications
Read 6 Reviews rated 4.3
using
NB400-105 in the following applications:

Publications
Read Publications using
NB400-105 in the following applications:

Reactivity Notes

Canine reactivity reported in scientific literature (PMID: 24612239)

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.05% Sodium Azide
Concentration
1 mg/ml
Purity
Immunogen affinity purified

Alternate Names for ABCA1 Antibody

  • ABC1
  • ABC-1
  • ABC1ATP-binding cassette transporter A1
  • ABCA1
  • ATP-binding cassette 1
  • ATP-binding cassette transporter 1
  • ATP-binding cassette, sub-family A (ABC1), member 1
  • CERP
  • Cholesterol efflux regulatory protein
  • EC 2.7.7.8
  • EC 2.8.1.8
  • EC 3.6.3
  • EC 3.6.3.41
  • FLJ14958
  • HDLDT1
  • membrane-bound
  • MGC164864
  • MGC165011
  • TGD
  • TGDATP-binding cassette sub-family A member 1

Background

ABCA1 (ATP-binding cassette transporter A1), ATP-binding cassette transporter family member, is a cAMP-dependent and sulfonylurea-sensitive anion transporter which is involved in the regulation of apolipoprotein AI (apoAI)-mediated cholesterol efflux from macrophages and act as a key gatekeeper influencing intracellular cholesterol transport. ABCA1 is widely expressed with highest expression levels found in macrophages and is induced by bacterial LPS which regulates its expression through LXR (liver X receptor) -independent mechanism. ABCA1 interacts with MEGF10 and its expression is repressed by ZNF202. ABCA1 phosphorylation at Ser-2054 site regulates the phospholipid efflux and DHHC8 mediated palmitoylation of ABCA1 is essential for its membrane localization. ABCA1 expressed in brain neurons/glia mediates cholesterol transport to apoE which is the major CNS apolipoprotein, and it influences the motor function and synaptic morphology also. Defects in ABCA1 have been linked to HDLD1 (high density lipoprotein deficiency type 1; also known as analphalipoproteinemia, Tangier disease or TGD) and HDLD2 (high density lipoprotein deficiency type 2; also known as familial hypoalphalipoproteinemia or FHA).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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AF7197
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NB400-105
Species: Hu, Mu, Rt, Po, Ca, Ch, Ha, Md
Applications: WB, Simple Western, ChIP, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP, GS

Publications for ABCA1 Antibody (NB400-105)(269)

We have publications tested in 10 confirmed species: Human, Mouse, Rat, Canine, Chicken, Chinese Hamster, Hamster, Mustelid, Porcine, Rabbit.

We have publications tested in 12 applications: ChIP, ELISA, EMSA, FLOW, IB, ICC/IF, IHC, IHC-FR, IHC-Fr, IHC-P, IP, WB.


Filter By Application
ChIP
(1)
ELISA
(1)
EMSA
(1)
FLOW
(1)
IB
(1)
ICC/IF
(17)
IHC
(11)
IHC-FR
(1)
IHC-Fr
(1)
IHC-P
(6)
IP
(8)
WB
(132)
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Filter By Species
Human
(66)
Mouse
(78)
Rat
(14)
Canine
(2)
Chicken
(1)
Chinese Hamster
(1)
Hamster
(6)
Mustelid
(1)
Porcine
(2)
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(1)
All Species
Showing Publications 1 - 10 of 269. Show All 269 Publications.
Publications using NB400-105 Applications Species
Zinkhan EK, Yu B, Schlegel A. Prenatal Exposure to a Maternal High Fat Diet Increases Hepatic Cholesterol Accumulation in Intrauterine Growth Restricted Rats in Part Through MicroRNA-122 Inhibition of Cyp7a1 Front Physiol 43249 [PMID: 29896121] (WB, Rat) WB Rat
Wang D, Hiebl V, Ladurner A et al. 6-Dihydroparadol, a Ginger Constituent, Enhances Cholesterol Efflux from THP-1-derived Macrophages Mol Nutr Food Res May 26 2018 [PMID: 29802792] (WB, Human) WB Human
Feng T, Liu P, Wang X et al. SIRT1 activator E1231 protects from experimental atherosclerosis and lowers plasma cholesterol and triglycerides by enhancing ABCA1 expression Atherosclerosis Apr 30 2018 [PMID: 29787963] (WB, Mouse) WB Mouse
Ishigami M, OgaSawara F, Nagao K et al. Temporary sequestration of cholesterol and phosphatidylcholine within extracellular domains of ABCA1 during nascent HDL generation. Sci Rep Apr 18 2018 [PMID: 29670126] (WB, Human) WB Human
Xie Z, Wang X, Liu X et al. Adipose-Derived Exosomes Exert Proatherogenic Effects by Regulating Macrophage Foam Cell Formation and Polarization. J Am Heart Assoc. Mar 3 2018 [PMID: 29502100] (WB, Mouse) WB Mouse
Casali BT, Reed-Geaghan EG, Landreth GE. Nuclear receptor agonist-driven modification of inflammation and amyloid pathology enhances and sustains cognitive improvements in a mouse model of Alzheimer's disease. J Neuroinflammation. 2018 Feb 15 [PMID: 29448961] (WB, Mouse) WB Mouse
Sallam T, Jones M, Thomas BJ, Wu X. Transcriptional regulation of macrophage cholesterol efflux and atherogenesis by a long noncoding RNA. Nat. Med. 2018 Feb 12 [PMID: 29431742] (WB) WB
Widenmaier SB, Snyder NA, Nguyen TB et al. NRF1 Is an ER Membrane Sensor that Is Central to Cholesterol Homeostasis Cell. 2017 Nov 16 [PMID: 29149604] (WB) WB
Doran AC, Ozcan L, Cai B et al. CAMKIIg suppresses an efferocytosis pathway in macrophages and promotes atherosclerotic plaque necrosis. J. Clin. Invest. 2017 Nov 01 [PMID: 28972541] (WB) WB
Storti F, Raphael G, Griesser V et al. Regulated efflux of photoreceptor outer segment-derived cholesterol by human RPE cells Exp. Eye Res. 2017 Sep 21 [PMID: 28943268] (ICC/IF, Human) ICC/IF Human
Show All 269 Publications.

Reviews for ABCA1 Antibody (NB400-105) (6) 4.36

Average Rating: 4.3
(Based on 6 reviews)
We have 6 reviews tested in 4 species: Human, Mouse, Rat, Other.

Reviews using NB400-105:
Filter by Applications
WB
(5)
IF
(1)
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Filter by Species
Human
(1)
Mouse
(3)
Rat
(1)
Other
(1)
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Images Ratings Applications Species Date Details
Western Blot ABCA1 NB400-105
Enlarge
reviewed by:
Jahangir Iqbal
WB Mouse 11/16/2015
View

Summary

ApplicationWestern Blot
Sample Testedmouse liver
SpeciesMouse
LotAF-2

Blocking

Blocking DetailsMembranes were blocked in 5% BSA in TBST for 1 h at room temperature

Primary Anitbody

Dilution RatioMembranes were immunoblotted with anti-ABCA1 antibody 1:2000 dilution in 2.5% BSA in TBST overnight at 4 oC

Secondary Antibody

Secondary DescriptionMembranes were incubated with secondary antibody conjugated to stabilized peroxidase for 1 hour at room temperature
Secondary Manufacturer Cat#Thermo Scientific # 32460
Secondary Concentrationdilution 1:15,000

Details

Detection NotesBands were visualized used chemiluminescence according to the manufacturer’s instructions

Comments

CommentsSamples were not boiled and 2X BME was added.
 
reviewed by:
Anonymous
WB Mouse 12/23/2014
View

Summary

ApplicationWestern Blot
Sample TestedSee PMID 23519696
SpeciesMouse

Blocking

Blocking DetailsSee PMID 23519696

Details

Detection NotesSee PMID 23519696

Comments

CommentsIndependent review from antibodyresource.comPublished in PMID 23519696
 
reviewed by:
Angeliki Chroni
IF Human 11/03/2014
View

Summary

ApplicationImmunofluorescence
Sample TestedSK-N-SH human neuroblastoma cells
SpeciesHuman
 
reviewed by:
Anonymous
WB Rat 10/15/2012
View

Summary

ApplicationWestern Blot
Sample TestedCardiomyocytes from newborn rats
SpeciesRat

Blocking

Blocking DetailsPBS Milk 5%, 1 hour, room temperature

Primary Anitbody

Dilution Ratio1000 = dilution, 1 hour, room temperature, PBS without tween and 2% Milk

Secondary Antibody

Secondary Descriptiongoat anti rabbit IgG HRP
Secondary Manufacturer Cat#Amersham NA934V
Secondary Concentration3000 diluted

Details

Detection NotesECL super plus Amersham RPN2132
 
reviewed by:
Cynthia Hong
WB Mouse 09/27/2011
View

Summary

ApplicationWestern Blot
Sample TestedMouse
SpeciesMouse
 
reviewed by:
Anonymous
WB Other 06/15/2009
View

Summary

ApplicationWestern Blot
Sample TestedRat astrocyte, Human Astrocyte, and Raw Mouse Neuron, Sample Amount: 25ug to 50ug
SpeciesOther

Blocking

Blocking DetailsBlocking Buffer: 5% Milk w/ TBST, Blocking Time: 1 hour, Blocking Temp: room temperature

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1/1000, Primary Ab Incubation Time: 3 hours, Primary Ab Incubation Temp: room temperature

Secondary Antibody

Secondary DescriptionSecondary Ab: anti-Rabbit IgG HRP, Secondary Ab Dilution Ratio: 1/1000

Details

Detection NotesExposure Time: 1 hour, Positive Control: HC/RA, Specific Bands: 200 kDa

Product General Protocols

Video Protocols

WB Video Protocol
ChIP Video Protocol
ChIP Webinar
ICC/IF Video Protocol

FAQs for ABCA1 Antibody (NB400-105). (Showing 1 - 10 of 11 FAQs).

  1. Do you have a protocol for Flow cytometry analysis with this ABCA1 antibody (NB400-105)?
    • Please see this link to our protocol for Flow cytometry. This is a general protocol that our lab uses as a guideline.
  2. We purchased this antibody a few weeks ago, and we by mistake stored it at -20 degrees Celsius. Is there possible the antibody still works?
    • Most likely the antibody should still work, but we cannot guarantee it. The biggest thing with antibody storage in most cases, especially for unconjugated antibodies, will be to avoid freeze/thaw cycles as much as possible. So even with ones that we recommend to freeze long term we still recommend aliquoting so they can go through as few of those as possible. I would say since it will probably only undergo one when you move it back to 4 degrees Celsius you should be fine, but again I can't say with 100% certainty. If you have a problem with detection using this product, please let us know so that we can figure out other possibilities to help you in that situation.
  3. I am interested in looking at this marker on monocytes by flow cytometry. I noticed that a directly conjugated 488 ab is available, and that your flow cytometry protocol calls for methanol fixation of the cells. Is the fixation required for antibody binding, or may it be used on cells that have not been methanol fixed? I would like to retain other markers.
    • Methanol is not required for antibody binding, but it is a fixation method that we found to not interfere with the antibody-antigen binding. You may use unfixed cells, but you may not be able to achieve staining. If you fix your cells with a formaldehyde-based fixative (we recommend antigen retrieval for IHC). However, as an inter-membrane protein, you will need to permeabilize your cells in order for the antibody to access the protein.
  4. To do Western blot for the determination of ABCA1 protein level, do I need to order NB400-105 or NB400-105B? Which one do you recommend if we also order the secondary antibody catalog # NB730-H?
    • The difference between NB400-105 or NB400-105B is biotin conjugation. It is up to you whether you would like to order the primary antibody that is conjugated or not. With the biotin-conjugated antibody (catalog # NB400-105B), you won't need a secondary, just a (strept)avidin-labelled detection reagent, such as streptavidin-HRP. With the unconjugated primary (catalog # NB400-105), you will need to use a secondary antibody. Some people say that when you use the secondary, there is an amplification of signal, because more than one secondary can bind to each primary. However, with the directly conjugated antibody you have fewer reagents and fewer wash steps.
  5. We purchased NB400-105, Lot# Z5, but the datasheet (approx. 450kDa) was differ from your website (approx. 220kDa) for size of WB. Which molecular weight is correct?
    • ABCA1 is highly glycosylated and has a predicted molecular weight of approximately 250 kDa without post-translational modification. The apparent molecular weight will change based on treatment and sample. All of the images on our website and datasheet are correct.
  6. Does this product contain any BSA?
    • No, this antibody does not have any BSA in its formulation.
  7. I bought ABCA1 antibody (NB400-105) from your company recently and used it in Flow cytometry. I want to know the binding site of ABCA1 and the antibody; is it within the cell or outside? Also, do I need to change the cytomembrane's permeabilization before my test?
    • The immuogen region of NB400-105 is within residues 1100-1300 of human ABCA1 and as a result it does not cover the extracellular domains, as a consequence, permeabilization is required for this antibody. As a general guideline, we usually recommend using 0.5-1.0% Triton detergent in the buffers in order to allow full permeabilization of our antibody.
  8. Is post-nuclear lysate is necessary? Can't I lysate my cells with RIPAbuffer (#R0278 sigma)?
    • The post-nuclear lysate you mentioned is posted on our website to demonstrate how we do the experiments, and how it was done by our collaborator (Ref Link #1). The procedure of getting the post-nuclear lysate was just for removing the nuclei before lysis and therefore the cleaner the lysates would be. You do not need to follow the method for the experiment that led to the result.
  9. Why shouldn't I boil the samples? is it OK to boil them if using RIPAbuffer?
    • To fractionate a membrane-bound protein, the lipid moieties associated with the protein frequently cause the smearing or streaky appearance of our WB. To result this difficulty, it is advised that we resuspend cell pellets in the SDS-sample buffer, left at room temp without boiling, spinning down the viscous cell debris, and then subject to PAGE and blotting. And this was the reason we suggested you not to boil your protein samples. Whether it will work for resuspending cell pellets in RIPA buffer, unfortunately we have no such data.
  10. Can i use DTT instead of Beta- mercaptoethanol?
    • Either DTT or 2-mercaptoethanol is routinely used in SDS-PAGE sample buffer (reducing gel electrophoresis). Except that 2-mercaptoethanol is a stronger reducing agent and is mostly frequently used for that purpose.
  11. Show All 11 FAQs.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB400-105AF405
Alexa Fluor 488 Labeled NB400-105AF488
Alexa Fluor 647 Labeled NB400-105AF647
Alexa Fluor 700 Labeled NB400-105AF700
Biotin Labeled NB400-105B
DyLight 405 Labeled NB400-105V
DyLight 488 Labeled NB400-105G
DyLight 550 Labeled NB400-105R
DyLight 650 Labeled NB400-105C
FITC Labeled NB400-105F
HRP Labeled NB400-105H
PE Labeled NB400-105PE

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Bioinformatics Tool for ABCA1 Antibody (NB400-105)

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Blogs on ABCA1. Showing 1-10 of 12 blog posts - Show all blog posts.

Niemann Pick-C1 and cholesterol dynamics
Niemann-Pick type C1 (NPC1) mediates low-density cholesterol transport from late endosomes and lysosomes to other areas of the cell via receptor mediation endocytosis.  Although cholesterol moves freely inside the cell, it cannot independently expo...  Read full blog post.

ABCA1 (ATP-binding cassette transporter A1)
The ABCA1 molecule is a primary gatekeeper for regulating the intracellular transport of cholesterol. It belongs to a larger related multifamily of cAMP-dependent anion transporter cell membrane molecules. These key proteins are responsible for tr...  Read full blog post.

ABCA1 - The Caretaker for Cholesterol Transportation
The ATP-binding cassette transporter A1 (ABCA1) protein is a key gatekeeper for regulating intracellular cholesterol transport. It is one member of a large family of genes comprised of cAMP-dependent anion transporter cell membrane proteins. These imp...  Read full blog post.

ABCG8: Cholesterol's Fate
The ATP-binding cassette (ABC) transporter genes are key gatekeeper molecules that regulate the amount of dietary cholesterol retained by the body. They are a multifamily comprised of cAMP-dependent anion transporter cell membrane proteins that monito...  Read full blog post.

ABCA1
ABCA1 is a key gatekeeper influencing intracellular cholesterol transport, and is an important member of a multifamily of cAMP-dependent anion transporter cell membrane proteins that regulate reverse cholesterol efflux from cells in peripheral tissues...  Read full blog post.

SREBP: Gatekeeper of Cholesterol Homeostasis
SREBP1 (sterol-regulatory-element-binding protein 2) is a basic-helix-loop-helix-leucine zipper (bHLH-ZIP) transcription factor. It regulates sterol and cholesterol homeostasis by controlling enzymes involved in cholesterol synthesis and uptake, e.g....  Read full blog post.

SREBP2: From Cholesterol Homeostasis to Cancer Invasion
Sterol-regulatory-element-binding protein 2 (SREBP2) is a transcription factor that regulates cholesterol homeostasis by controlling enzymes involved in cholesterol synthesis and uptake, e.g. HMG-CoA. Along with another transcription factor LXR, SREBP...  Read full blog post.

LXR Alpha, ABCA1 and Cholesterol Homeostasis
LXR Alpha, also known as Liver X receptor Alpha is a 50KDa protein that belongs to the nuclear hormone receptor family located in the nucleus. It is specifically expressed in the liver, kidney and intestine; however it has also been found in the splee...  Read full blog post.

The ABCA1 Antibody and ABC Transporter Research
The ABC (ATP-binding cassette) transporters are a superfamily of transmembrane proteins which play an essential role in membrane transport in both prokaryotes and eukaryotes. In humans, ABC transporters have been implicated in many conditions. For exa...  Read full blog post.

Exploring how ABCG8 Affects Heart Disease
The high prevalence of atherosclerosis in developed nations is not breaking news; it has been well established that heart disease is the leading cause of death in the United States, and it presents a major socioeconomic burden. Investigators across th...  Read full blog post.

Showing 1-10 of 12 blog posts - Show all blog posts.

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Recent Reviews

4.3
5
3
4
2
3
1
2
0
1
0

Jahangir Iqbal
11/16/2015
Application: WB
Species: Mouse

 
Anonymous
12/23/2014
Application: WB
Species: Mouse

 
Angeliki Chroni
11/03/2014
Application: IF
Species: Human

Bioinformatics

Gene Symbol ABCA1
Entrez
Uniprot