Overview

  • Product name
    Anti-Scavenger Receptor BI + BII antibody
  • Description
    Rabbit polyclonal to Scavenger Receptor BI + BII
  • Host species
    Rabbit
  • Specificity
    ab36970 is specific for SR-BI and SR-BII.
  • Tested applications
    Suitable for: WB, ICC/IF, IP, Blockingmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide, corresponding to extracellular amino acids 230-280 of Scavenger Receptor BI + BII

  • Positive control
    • Tissues that express SR-BI and/or SR-BII such as such as liver, ovary, adrenal glands, and to as lesser extent testes, heart and mammary glands.

Applications

Our Abpromise guarantee covers the use of ab36970 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
WB 1/1000. Predicted molecular weight: 82 kDa.
ICC/IF 1/1000.
IP 1/100.
Blocking 1/100.

Target

  • Relevance
    High density lipoproteins (HDLs) play a critical role in cholesterol metabolism and their plasma concentrations are inversely correlated with risk for atherosclerosis. SR-BI and SR-BII (previously known as SR-BI.2) are the alternatively spliced products of a single gene. SR-BII and SR-BI are identical except for the encoded c-terminal cytoplasmic domain. Both SR-BI and SR-BII bind HDL and mediates selective uptake of HDL cholesteryl ester, but with SR-BII having an approximately 4-fold lower efficiency than SR-BI. SR-BI and SR-BII are expressed primarily in liver and non-placental steroidgenic tissues. Although the role of these scavenger receptors is not completely clear, SR-BII mRNA results from the alternative splicing of SR-BI precursor transcripts with both isoforms mediating selective transfer of lipid between HDL and cells. Therefore, the relative expression and functional activities of these two isoforms create a potential means of regulating selective lipid transfer between HDL and cells.
  • Cellular localization
    Lysosome; lysosomal membrane; multi-pass membrane protein.
  • Database links
  • Alternative names
    • CD36 and LIMPII Analogous 1 antibody
    • CD36L1 antibody
    • CD36L2 antibody
    • CLA1 antibody
    • HLGP85 antibody
    • LGP85 antibody
    • LIMP II antibody
    • SCARB1 antibody
    • SCARB2 antibody
    • Scavenger receptor class B member 2 antibody
    • Scavenger Receptor Class B Type I antibody
    • SRBI antibody
    • SRBII antibody
    see all

Images

  • All lanes : Anti-Scavenger Receptor BI + BII antibody (ab36970) at 1/1000 dilution

    Lane 1 : wild-type mice
    Lane 2 : SR-BI deficient mice

    Predicted band size: 82 kDa

  • Anti-Scavenger Receptor BI + BII antibody (ab36970) at 1/1000 dilution + 80 µg of total mouse liver lysates

    Predicted band size: 82 kDa

References

This product has been referenced in:
  • Balasubramanian K  et al. Dichotomous roles for externalized cardiolipin in extracellular signaling: Promotion of phagocytosis and attenuation of innate immunity. Sci Signal 8:ra95 (2015). Read more (PubMed: 26396268) »
  • Badeau RM  et al. Human macrophage cholesterol efflux potential is enhanced by HDL-associated 17beta-estradiol fatty acyl esters. J Steroid Biochem Mol Biol 116:44-9 (2009). Blocking ; Human . Read more (PubMed: 19406243) »
See all 3 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Answer

Thank you for taking the time to contact us. I am sorry to hear you have some concerns regarding this antibody.

In this case, I would suggest the white powder you are seeing may be precipitate. We recommend that aliquots of antibody should be no smaller than 10 µl where possible. The smaller the aliquot, the more the stock concentration is affected by evaporation and adsorption of the antibody onto the surface of the storage vial. This evaporation could also result in precipitation, particularly if the antibody is stored frozen for a long period, such as a year.

I can recommend to mix the precipitate very well using a pipette, and try vortexing as well. I hope this would provide a more homogenous solution for you to try in your experiments.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for WB, ICC/IF, IP, blocking and in mouse and human samples. We can also guarantee the general quality of the antibody for 6 months. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund. In this case, I am sorry to confirm that as the antibody was purchased over 6 month ago it will regrettably no longer be covered by our 6 month guarantee on this occasion.

Please do not hesitate to let me know if you have any further questions or concerns.

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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.

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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.

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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.

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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.

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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.

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Answer

Thank you for your enquiry

We obtain this product from an external source and unfortunately they are not able to provide specific information about the sample preparation.

I reviewed the literature and found the following protocol that you may find useful:

1. Used 1-2 liver lobes.
2. Homogenize tissue in PBS/Protease Inhibitor cocktail.
3. After centrifugation resuspend the pellet in approx. 400µl RIPA lysis buffer 30min (vortex every 5-10 min).
4. Centrifuge to give whole cell extracts.
5. To improve the amount of sample extracted, repeat this lysis step with the pellet from the centrifugation and pool the supernatant of the 1st and the 2nd lysis.

We also stock a range of liver lysates:

Liver (Mouse) Whole Cell Lysate - normal tissue, 0 days old
www.abcam.com/ab7264

Liver (Mouse) Whole Cell Lysate - normal tissue, 7 days old
www.abcam.com/ab7266

Liver (Mouse) Whole Cell Lysate - normal tissue
www.abcam.com/ab29301

Liver (Human) Tissue Lysate - adult normal tissue
www.abcam.com/ab29889

Please let me know if there is any more information you need.

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