Overview

  • Product name

  • Description

    Rabbit polyclonal to ABCA1
  • Host species

    Rabbit
  • Specificity

    ab7360 has previously been shown to work well in ICC/IF on HeLa cells. However recent batches of this antibody, do not work in this application. We would recommend ab18180 for researchers wanting to detect ABCA1 in ICC/IF. For further information, please contact our Scientific Support Team.
  • Tested applications

    Suitable for: IP, ELISA, WB, IHC-P, IHC-Frmore details
  • Species reactivity

    Reacts with: Mouse, Human, Monkey
  • Immunogen

    Synthetic peptide corresponding to Human ABCA1. Synthetic peptide: The immunogen is generated from within residues 1100-1300 of Human ABCA1
    Database link: O95477

Applications

Our Abpromise guarantee covers the use of ab7360 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
IP Use at an assay dependent concentration.

PMID: 14660648

ELISA Use at an assay dependent concentration.

PMID: 21829447

WB 1/500 - 1/1000. Predicted molecular weight: 220 kDa. Additional non-specific bands are seen at lower molecular weights, but do not interfere with the ABC1 signal. It is important not to boil the sample before loading onto the gel. Boiling can cause aggregation in large proteins, resulting in the proteins inability to enter into the gel.
IHC-P 1/500.
IHC-Fr 1/750. PubMed: 12547833

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

  • 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

  • All lanes : Anti-ABCA1 antibody (ab7360)

    Lane 1 : RAW 264.7 whole cell lysate treated with T0901317
    Lane 2 : RAW 264.7 whole cell lysate untreated

    Lysates/proteins at 25 µg per lane.

    Predicted band size: 220 kDa

  • ab7360, staining ABCA1 in prostate epithelium showing luminal and membrane staining by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
  • The experiment was performed by treating RAW macrophages with 9-cis-retinoic acid and 22R-hydroxycholesterol, known inducers of ABCA1expression in macrophages. Then total cell post-nuclear lysate (40ug protein) was separated by SDS-PAGE and detected using a 1:1000 dilution of ab7360 affinity purified Lot G incubated for 1 hour at room temperature (Lane A). Although there are lower molecular weight bands on the blot, the ABCA1 signal is excellent and gives the expected 3 bands. It is not known why ABCA1 runs as three bands, but it has been found to do so by many researchers. It is probably due to protein modifications such as glycosylation. The antibody was also tested against ABCA1 transiently expressed in 293 cells as an independent test with excellent results.

    The experiment was performed by treating RAW macrophages with 9-cis-retinoic acid and 22R-hydroxycholesterol, known inducers of ABCA1expression in macrophages. Then total cell post-nuclear lysate (40ug protein) was separ
  • Detection of ABCA1 in mouse peritoneal macrophages using ab7360. ECL exposure, 1 min.
    Lane 4: T09 uninduced lysate
    Lane 5: T09 induced lysate Detection of ABCA1 in mouse peritoneal macrophages using ab7360. ECL exposure, 1 min.
    Lane 4: T09 uninduced lysate
    Lane 5: T09 induced lysate

References

This product has been referenced in:

  • Takeuchi S  et al. Elevated Membrane Cholesterol Disrupts Lysosomal Degradation to Induce ß-Amyloid Accumulation: The Potential Mechanism Underlying Augmentation of ß-Amyloid Pathology by Type 2 Diabetes Mellitus. Am J Pathol 189:391-404 (2019). Read more (PubMed: 30448407) »
  • Liu JY  et al. Protective effect of down-regulated microRNA-27a mediating high thoracic epidural block on myocardial ischemia-reperfusion injury in mice through regulating ABCA1 and NF-?B signaling pathway. Biomed Pharmacother 112:108606 (2019). Read more (PubMed: 30802823) »
See all 45 Publications for this product

Customer reviews and Q&As

11-20 of 21 Abreviews or Q&A

Application
Western blot
Sample
Human Tissue lysate - other (placenta, arteries)
Loading amount
50 µg
Specification
placenta, arteries
Treatment
50U Benzonuclease for 1 h
Blocking step
Milk as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 7

Abcam user community

Verified customer

Submitted Aug 23 2006

Answer

Thank you for your reply and the details of the experiment. You do not seem to mention any blocking agent in the experiment and we have found that it is important to block the membrane in 5% milk for 1hr at RT, washing in TBST and adding the antibody diluted in 3% milk in TBST. We have noted on the datasheet that "Additional non-specific bands are seen at lower molecular weights, but do not interfere with the ABC1 signal" but if you still do not see a band at the correct MW with a blocking step please do not hesitate to contact me, providing your order details and I will arrange a replacement of the other antibody if the antibody was purchased in the last 90 days,

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Answer

I'm very sorry to hear you have been experiencing problems with ab7360. The fact that you are having good results with ab18180 rules out that the problem is due to your samples, and I would appreciate if you could tell me more about the conditions you have used ab7360 in, as one or more will be responsible for the problem, or the antibody is damaged. -what dilution have you tested? -what incubation time have you tested? -what blocking buffer have you used? -what antibody dilution buffer have you used? -have you tried the secondary antibody alone; or with other primary antibodies? -how did you store the antibody? If you could please provide your order details I will also look at possible shipping delays which would indicate a damaged vial. If the problem is due to the antibody and it was purchased in the last 90 days I can certainly offer you a replacement vial of ab18180. I look forward to hearing from you to resolve this matter, thank you in advance for those details,

Read More

Answer

Thank you for your enquiry. To our knowledge, this antibody has not yet been tested for application in ELISA. It doesn't mean that the antibody won't work, we just don't know. We have some very nice ELISA protocols available on our website which should be helpful for you. To find these protocols, you can click on the "protocols" tab located on the online product or by clicking on the "resources" menu located on the Abcam homepage. Please let us know how you get on by submitting an Abreview and in return we will award you 50 Abcam Points, which can be redeemed on a number of rewards (a further 100 Abcam Points will be offered for an image). Please contact us again if you have any additional questions.

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Answer

We do not have an exact protocol but I do have the following information that should provide useful: Postnuclear samples means that the cells are broken open either by sonication or dounce homogenization and the nclei are removed by low speed centrifugation. The resulting supernatant is called postnuclear. This supernatant contains most of the cytosolic and membrane proteins without containing nuclear components. Most of the references listed on the datasheet(that are doing westerns) list a brief description of how they obtained postnuclear lysates but it appears that none are highly detailed. I hope this helps.

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Answer

I'm pleased to hear that this antibody is working well for you. Ab7360 is not affinity-purified; the available purity is whole antiserum. Unfortunately, we do not have blocking peptide or a specific positive control lysate available for ab7360. The batch number should be on the vial-for example, G6 (27409)or 29826. Do you have a number such as that on your vial? Please confirm your batch number and I can check to see if we still have that batch in stock.

Read More

Answer

Here is the procedure with a typical blot and important information found here (http://www.novus-biologicals.com/abc/NB400-105.html): 1. Load 40 ug postnuclear lysates to 7.5% or 4-15% Tris-HDL SDS gel (Bio-RAD) in sample buffer without boiling. 2. Transfer to nitrocellulose membrane at 100V 1hr or 30V overnight. 3. Block membrane in 5% milk in TBS-T for at least 1 hr. Wash with TBS-T 5 minutes. 4. Blot with anti-ABCA1 antibody in 3% milk in TBS-T for 1 hour. 5. Wash with TBS-T 3 times, 10 minutes each. 6. Blot with donkey anti-rabbit 2' antibody (Amersham) at 1:2000 in 3% milk in TBS-T for 1 hour. 7. Wash with TBS-T 3 times, 10 minutes each. 8. Detect with chemiluminescent reagent (Pierce) 9. Expose to X-ray film, 30sec, 1min, and 3 min. TBS-T: Tris-buffered-saline with tween 20. Specific References: http://www.novus-biologicals.com/ProdRefPages/pr400-105.html

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Answer

The ABCA1 antibody may work, but I suspect the background would be too high since this antibody was made in rabbit.

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Answer

Our ABCA1 antibody, NB 400-105, does not work well (if at all) for IH. As far as we know, it has only been used for western analysis.

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Answer

Dear Sir: The source of protein for our assay was induced RAW macrophages. Most cell types express low levels of ABC1. Therefore,expression needs to be induced by using 22-hydroxycholesterol and 9-cis-retonoic acid as ligands for the transcription factor LXR. I hope this information is helpful. Best Regards, Kate Lynott

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11-20 of 21 Abreviews or Q&A

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