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:
  • Zhang X  et al. Human Gingiva-Derived Mesenchymal Stem Cells Modulate Monocytes/Macrophages and Alleviate Atherosclerosis. Front Immunol 9:878 (2018). Read more (PubMed: 29760701) »
  • Zhao ZW  et al. Heat shock protein 70 accelerates atherosclerosis by downregulating the expression of ABCA1 and ABCG1 through the JNK/Elk-1 pathway. Biochim Biophys Acta 1863:806-822 (2018). WB . Read more (PubMed: 29678642) »
See all 37 Publications for this product

Customer reviews and Q&As

1-10 of 21 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa and pluripotent cells)
Permeabilization
Yes - Triton X-100
Specification
HeLa and pluripotent cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 14 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (SCC61 cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
40 µg
Specification
SCC61 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Nov 13 2015

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued an alternative free of charge replacement of ab7360 with the order number 1214026.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Question

Dear Tineke/Abcam,

We purchased the following Abcam antibodies from Sapphire Biosciences as one single order which were shipped to us as a single lot:

1 AB68466 Rabbit polyclonal to PSAP

1 AB36970 Rabbit polyclonal to Scavenger Receptor BI + BII

1 AB7360 Rabbit polyclonal to ABCA1

1 AB3366 Mouse monoclonal [JSB-1] to P Glycoprotein

1 AB117778 Rabbit polyclonal to Cingulin

1 AB72978 Mouse polyclonal to CNKSR3 Based on our published work these 4 proteins should be expressed in differentiated Caco-2 cells

1 AB111692 Rabbit polyclonal to MAGI3

1 AB83891 Rabbit polyclonal to MPP5


The only one that has worked satisfactorily is AB68466 Rabbit polyclonal to PSAP (see attached file showing these blots). All the others have failed to work as promised. Please note, as previously mentioned, we have used a variety of exposure times with our blots – we have routinely used 10 different exposure times (as the signal increased and also when the signal would have been waning – depending on the results) for each blot using your antibodies to try and optimise what we can visualise with our blots. This has come at some considerable expense in terms of time, developing chemicals and film.

Yes - the other antibodies we tested from our preferred supplier worked. These were purchased at the same time we purchased the Abcam antibodies but they use a different supplier in NZ so were shipped separately. They were tested at the same time we tested yours, using identical conditions and protein samples, and had very low background which is what we have come to expect from their antibodies.

In our previous emails, we filled in questionnaires associated with the failure of AB36970 Rabbit polyclonal to Scavenger Receptor BI + BII, AB7360 Rabbit polyclonal to ABCA1, AB3366 Mouse monoclonal [JSB-1] to P Glycoprotein, and AB117778 Rabbit polyclonal to Cingulin. Despite filling in your questionnaires, you have asked for additional information, which we provided, EXCEPT for the blots as this has taken some considerable time to organise. I have been away at a conference, and as I also work part-time, this has been a pain to organise for you.

During this time we were also evaluating the remaining 4 antibodies and have subsequently found these to be unsatisfactory too. Please see the attached questionnaires for these antibodies. This report of failure to deliver is also within the 6 month time frame of when we received your antibodies.

Do I need to point out, yet again, that we are not stupid. You talked about the samples we tested against (rat, bovine and human Caco-2 cells) and that the antibodies we purchased were not known to cross-react with bovine samples (except MPP5 which was predicted to cross-react). The reason we chose these samples is that although our first set of experiments will ultimately be performed using differentiated Caco-2 cells, some of these targets are highly expressed in liver and we wanted to see if bovine liver could be used as a control – which is why you will note in our blots that we always ran these next to Caco-2 cell extracts to see how they compared. The same goes for the rat colon/rat colon epithelial scrapings – most of the antibodies were known to cross-react with rat or mouse as well as human proteins (also ab36970 is listed as being predicted to cross-react with rat which is not what you said in the email below). Obviously we had good reasons why we wanted to this - we will be doing animal trials in the future amongst other reasons - and again WE ALWAYS COMPARED OUR RESULTS TO OUR HUMAN CACO-2 CELL RESULTS.

I estimate that we have probably spent more money on staff time and other lab consumables trying to get your antibodies to work and fill in your questionnaires, answer yet more questions from you, send you copies of our blots....etc....than it cost us to purchase the antibodies in the first place. This is the first time I have used Abcam antibodies as I usually purchase them from another supplier but they were unable to supply these particular antibodies. My expectation was that if your antibodies performed as expected, I would continue to purchase all my antibodies from you. Given my experience, I will not be purchasing antibodies from you in the future.

Please either replace these antibodies immediately, or better still, please refund our money in full so we can purchase them elsewhere. I don’t want to be bothered with any further questions. You have wasted enough of our time and it’s about time you did the right thing by us.

Read More
Answer

Thank you for your reply and all the additional information. This makes the problems you have been experiencing much clearer to understand. It is clear that you have spent a lot of time in trying to optimise these antibodies and I am sorry that they are not providing the expected results.

I am sorry if you feel the questions I have been asking have not been appropriate. The reason I askedfor further information is to try to understand what has been tried already and to try to understand what may be contributing to these problems. With a total of 7 antibodies not working as expected we are taking this very seriously to try and isolate the reason and to offer you a suitable solution as well as preventing this from happening again in the future.

I have been discussing this case with my colleague Dr. Bagrij who has experience in working with a number of the targets you are interested in. I understand the Caco2 lysate you have is very limited and I don't want you to use any more in optimising any of these antibodies. I think it would be worthwhile to try a different lysate to the ones you have been using to see if this makes a difference in the results observed. To this end I would like to send you a pre-prepared Jurkat lysate which have been used as positive controls for the anti-Cingulin antibody (ab117778) and the anti-MPP5 antibody (ab83891). Would you mind trying this lysate with these two antibodies just to see if the expected signal is observed?

You mentioned that you have previously published in regards to the Caco2 cells expressing a number of these targets. Would you mind sharing this reference with me? This information would be very useful in this investigation as well asfor other customers using these antibodies.

Could you also confirm on which date you received the 8 antibodies? I know which order they came from but I want to make sure they reached youwithin the expected time.Were they all delivered in the same package? What was the package like?

I am sorry that you are having these problemsand Iwould like to assure you that I amtrying my best to get to the bottom of this and provide you with a solution.

I look forward to receiving your reply.

Read More

Question

Please see below for the response from the customer;

I’m very frustrated by Abcam’s response. It took a long time to fill in the questionnaires for each of the antibodies and some of their additional questions were already answered in the questionnaire (Q3 and Q6). Q6 is quite insulting, even more so when I had specifically answered that I’d used anti-mouse secondary antibodies.

It also took them 4-5 working days to reply to the questionnaire answers, now they want more information, taking more of my time, and I guess I have to wait another 4-5 working days for their next response.

Quite frankly, this is unacceptable.

In short,

Q1. the same blocking agent was used consistently throughout the western experiment. So, if BSA was used for blocking, it was also used for the primary and the secondary. When it didn’t work, we tried milk etc.

Q2. For most of the repeats, blots weren’t stripped. We made ran fresh gels and blotted these (exactly the same loading concentrations and conditions). Therefore, stripping was not the issue. ALWAYS, we ran another gel with the same loading concentration which was stained with coomassie. ALWAYS we ran coloured markers so we could check that the protein had transferred. We have never had trouble with transfer using the i-blot.

Q3. Please read the answers I have already supplied.

Q4. Exposure times varied from 15 s through to 15 min. We also used 2 pieces of film so that the bottom film acted as a filter. We developed at least 8 pieces of film for each blot each time we repeated it and also used a range of exposure times. Faint blots were left longer while those with high background were just left for a few seconds. High background blots were also re-exposed after 1-2 hours as the signal was waning. We covered the entire range.

Q5. This will take a lot of time as we’ve done lots of blots. They will have to insist this is absolutely necessary then pay for my time. It’s their antibodies that aren’t working so I shouldn’t have to defend my results. My time will probably cost more than their antibodies do to replace.

Q6. The secondary antibodies were brand new and worked because, as previously answered, we had magic markers which only show up if the secondary antibody is working.

My expectation is that Abcam will hurry up and come to the party with new, replacement antibodies in a timely manner and stop wasting our time. If they don’t, we won’t be using them again and as we work for a large research institution with a large international network of collaborators, won’t be recommending them to anyone.

Read More
Answer

Thank you for your reply.

I am very sorry that you have not been satisfied with the way we have handeled your complaints. I do understand and appreciate the amount of time you have spent it the laboratory. Our policy is that we are happy to offer a refund, credit note or free of charge replacement when a product is not working in a successfully tested applications or species (and the product has been purchased in the last 180 days).

However, we often find that suggesting some scientifically thought out optimisation tips improves the results. This is obviously a preferable outcome.So I hope you can understand that our policy is to offer the best technical support possible to help optimise the results first.

It is also important for us to understand what is contributing to the problems encountered, as if there is a problem with the antibody, forexample with a particular lot, we need to try to find out why this has happened and if any other customers may be affected and if we need to let them know.

I am sure you can understand that with four antibodies not providing satisfactory results for you we need to take it especially seriously, I do not want to simply offer replacement for them to perform in exactly the same way and further waste your time.

I am sorry that I did not read the information regarding the anti-mouse antibody used with ab3366 properly. This was an error on my part and I apologize. The reason I wanted to have a look at the images of the blots was to try to ascertain where the high background may be coming from. With the antibodies used, none have been tested with bovine samples so we would not be able to know if they would cross react or not. Similarly, ab7360 and ab36970 have not been tested with rat samples as yet, although they would be predicted to cross-react due to their sequence homology. That leaves onlytheCaco-2 cell samples for ab7360 and ab36970. It would be very helpful to understand the results observed more fully if you would be able to provide a few representative images of the blots obtained.

You have mentioned that other antibodies have been used with the samples and have performed as expected. Are any of these from Abcam, or are they from other companies? Were any of them delivered in the same shipment as the 4 antibodies which are not working? I am trying to ascertain if there may have been a problem with your particular delivery which affected the antibody activity. This is important as with some of the antibodies involved, we only have the same lots available as you have already tried and I need to ascertain if this is worth trying again for you.

I am sorry for the delay youhave experienced in our handling of your complaints. I am sorry for this and the inconvenience caused. I will try to resolve the situation as quickly as I can.

Read More

Answer

Thank you for reporting the problems encountered with these antibodies and for taking the time to complete the questionnaire. I am sorry to hear that these antibodieshave not providing satisfactory results so far.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, there are a few further details I would like clarified if possible:

1. You have mentioned that BSA, milk and Roti block were tried. When each blocking agent was tried, was the same blocking used in the primary antibody diluent? For example, when using the BSA blokcing, did you use BSA in the antibody diluent or did you continue with milk for this? Was this the case for the secondary antibody as well?

2. Could you explain how the membranes were stripped between each staining?

3. Were the same transfer conditions used for each antibody?

4. What exposure times were used?

5. Could youshare some images of the blots obtained? With each lane labelled with the samples and the molecular weight markers. If its not too much trouble, could the images of the blotting with the xxxxxx antibody and the acting loading control also be included. This would help greatly in understanding more fully the problems encountered with the antibodies.

6. With the anti-P Glycoprotein antibody (ab3366), did you use the same secondary antibody as with the other antibodies? Please bear in mind that this is a mouse monoclonal antibody and will not be detected by the anti-rabbit secondary antibody. Was a different antibody used? If so, has this one been used successfully previously?

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I look forward to receiving your reply.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (human astrocyte)
Loading amount
25 µg
Specification
human astrocyte
Treatment
treated with LXR ligand for 20 hrs
Gel Running Conditions
Reduced Denaturing (4-15% gradient gel)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Aug 06 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293, THP1, mouse BMDM, mouse tissue)
Loading amount
15 µg
Specification
HEK293, THP1, mouse BMDM, mouse tissue
Treatment
Transfected with ABCA1 expression plasmid, Treat with 1uM TO-1317 for 24hr
Gel Running Conditions
Reduced Denaturing
Blocking step
5%milk+1%BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Jan 26 2008

Answer

I'm sorry to hear you are having a problem with ab7360. I would like to suggest the following modifications to your protocol: The recommended dilution range for this antibody is 1/500 - 1/1000. You may be using it at too high a dilution. I did not see your incubation conditions with this antibody from your enquiry. Since you are seeing no signal, I would suggest removing the milk from the primary antibody solution and incubating at 4C overnight. The milk in the primary antibody solution can overblock the membrane, preventing the antibody from binding to its target. I would continue incubating the sample in loading buffer for 30 minutes at 37C. This protein can form aggregates, preventing migration into the gel matrix. For how long did you block? If you have blocked for longer than one hour, you may wish to reduce this step to 1 hour, or even 30 minutes. Did you check (e.g. Ponceau stain) to see that the transfer was efficient? Please let me know if this helps and do not hesitate to contact us for further advice.

Read More
Application
Western blot
Sample
Mouse Tissue lysate - whole (macrophage cell line)
Loading amount
15 µg
Specification
macrophage cell line
Treatment
0.3mM dbcAMP 24 hr
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5

Abcam user community

Verified customer

Submitted Sep 18 2006

1-10 of 21 Abreviews or Q&A

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