Overview

  • Product name
    Anti-CD36 antibody [FA6-152]
    See all CD36 primary antibodies
  • Description
    Mouse monoclonal [FA6-152] to CD36
  • Host species
    Mouse
  • Tested applications
    Suitable for: Blocking, Functional Studies, IHC-Fr, Flow Cytmore details
    Unsuitable for: IHC-P or WB
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    20-weeks-old fetal erythrocytes.

  • Positive control
    • Flow cytometry: THP-1 cells. HEL cells
  • General notes

    We have data from customer abReviews and publications indicating that this antibody is capable of detecting CD36 via western blot. However, based on customer feedback of difficulties and extensive troubleshooting using this antibody in western blot, we are removing WB as a tested application and recommend ab133625 as an alternative for use in this application.

Applications

Our Abpromise guarantee covers the use of ab17044 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
Blocking Use at an assay dependent concentration. Blocks collagen/thrombospondin binding.
Functional Studies Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

  • Application notes
    Is unsuitable for IHC-P or WB.
  • Target

    • Function
      Multifunctional glycoprotein that acts as receptor for a broad range of ligands. Ligands can be of proteinaceous nature like thrombospondin, fibronectin, collagen or amyloid-beta as well as of lipidic nature such as oxidized low-density lipoprotein (oxLDL), anionic phospholipids, long-chain fatty acids and bacterial diacylated lipopeptides. They are generally multivalent and can therefore engage multiple receptors simultaneously, the resulting formation of CD36 clusters initiates signal transduction and internalization of receptor-ligand complexes. The dependency on coreceptor signaling is strongly ligand specific. Cellular responses to these ligands are involved in angiogenesis, inflammatory response, fatty acid metabolism, taste and dietary fat processing in the intestine (Probable). Binds long-chain fatty acids and facilitates their transport into cells, thus participating in muscle lipid utilization, adipose energy storage, and gut fat absorption (By similarity) (PubMed:18353783, PubMed:21610069). In the small intestine, plays a role in proximal absorption of dietary fatty acid and cholesterol for optimal chylomicron formation, possibly through the activation of MAPK1/3 (ERK1/2) signaling pathway (By similarity) (PubMed:18753675). Involved in oral fat perception and preferences (PubMed:22240721, PubMed:25822988). Detection into the tongue of long-chain fatty acids leads to a rapid and sustained rise in flux and protein content of pancreatobiliary secretions (By similarity). In taste receptor cells, mediates the induction of an increase in intracellulare calcium levels by long-chain fatty acids, leading to the activation of the gustatory neurons in the nucleus of the solitary tract (By similarity). Important factor in both ventromedial hypothalamus neuronal sensing of long-chain fatty acid and the regulation of energy and glucose homeostasis (By similarity). Receptor for thombospondins, THBS1 and THBS2, mediating their antiangiogenic effects (By similarity). As a coreceptor for TLR4:TLR6 heterodimer, promotes inflammation in monocytes/macrophages. Upon ligand binding, such as oxLDL or amyloid-beta 42, interacts with the heterodimer TLR4:TLR6, the complex is internalized and triggers inflammatory response, leading to NF-kappa-B-dependent production of CXCL1, CXCL2 and CCL9 cytokines, via MYD88 signaling pathway, and CCL5 cytokine, via TICAM1 signaling pathway, as well as IL1B secretion, through the priming and activation of the NLRP3 inflammasome (By similarity) (PubMed:20037584). Selective and nonredundant sensor of microbial diacylated lipopeptide that signal via TLR2:TLR6 heterodimer, this cluster triggers signaling from the cell surface, leading to the NF-kappa-B-dependent production of TNF, via MYD88 signaling pathway and subsequently is targeted to the Golgi in a lipid-raft dependent pathway (By similarity) (PubMed:16880211).
      (Microbial infection) Directly mediates cytoadherence of Plasmodium falciparum parasitized erythrocytes and the internalization of particles independently of TLR signaling.
    • Involvement in disease
      Platelet glycoprotein IV deficiency
      Coronary heart disease 7
    • Sequence similarities
      Belongs to the CD36 family.
    • Post-translational
      modifications
      N-glycosylated and O-glycosylated with a ratio of 2:1.
      Ubiquitinated at Lys-469 and Lys-472. Ubiquitination is induced by fatty acids such as oleic acid and leads to degradation by the proteasome (PubMed:21610069, PubMed:18353783). Ubiquitination and degradation are inhibited by insulin which blocks the effect of fatty acids (PubMed:18353783).
    • Cellular localization
      Cell membrane. Membrane raft. Golgi apparatus. Apical cell membrane. Upon ligand-binding, internalized through dynamin-dependent endocytosis.
    • Information by UniProt
    • Database links
    • Alternative names
      • Adipocyte membrane protein antibody
      • BDPLT10 antibody
      • CD36 antibody
      • CD36 antigen (collagen type I receptor, thrombospondin receptor) antibody
      • CD36 antigen antibody
      • CD36 molecule (thrombospondin receptor) antibody
      • CD36 molecule antibody
      • CD36_HUMAN antibody
      • CHDS7 antibody
      • Cluster determinant 36 antibody
      • Collagen receptor, platelet antibody
      • FAT antibody
      • Fatty acid translocase antibody
      • Fatty acid transport protein antibody
      • Glycoprotein IIIb antibody
      • GP IIIb antibody
      • GP3B antibody
      • GP4 antibody
      • GPIIIB antibody
      • GPIV antibody
      • Leukocyte differentiation antigen CD36 antibody
      • MGC108510 antibody
      • MGC91634 antibody
      • PAS 4 protein antibody
      • PAS IV antibody
      • PAS-4 antibody
      • PASIV antibody
      • Platelet collagen receptor antibody
      • Platelet glycoprotein 4 antibody
      • Platelet glycoprotein IV antibody
      • scarb3 antibody
      • Scavenger receptor class B member 3 antibody
      • Thrombospondin receptor antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) of human breast carcinoma stained for CD36 with ab17044 at 20 µg/ml

    • Detection of CD36 in THP-1 cells. Red, black and blue line represent the isotype control, cells only and ab17044 at 10 μg/ml, respectively.

    • Overlay histogram showing THP1 cells stained with ab17044 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab17044, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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 result in 80% methanol (5 min) fixed THP1 cells used under the same conditions.

      Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

    References

    This product has been referenced in:
    • Alva-Murillo N  et al. Sodium Octanoate Modulates the Innate Immune Response of Bovine Mammary Epithelial Cells through the TLR2/P38/JNK/ERK1/2 Pathway: Implications during Staphylococcus aureus Internalization. Front Cell Infect Microbiol 7:78 (2017). Read more (PubMed: 28361042) »
    • Lopez-Carmona MD  et al. CD36 overexpression: a possible etiopathogenic mechanism of atherosclerosis in patients with prediabetes and diabetes. Diabetol Metab Syndr 9:55 (2017). Read more (PubMed: 28729885) »
    See all 31 Publications for this product

    Customer reviews and Q&As

    1-10 of 23 Abreviews or Q&A

    Question

    Dear Abcam



    I have previously used a CD36 antibody from you (ab36977) with success but have noticed that it unfortunately no longer is available. Instead we tried ab78054 (with major problems, which we had a mail correspondence with you about in November 2011) and now we have tried the ab17044 but unfortunately also with problems.



    The homogenate used for testing the ab17044 has also been used for the other two antibodies :

    Human skeletal muscle tissue homogenized in Tris buffer (25mM Tris, 5mM EDTA, 3% SDS including Protease and phosphatase inhibitors: 20mM β-glycerophosphate, 10mM pyrophosphate,2mM NaOrtovanadate, 2.5mM PMSF, 1 mini comple protease inhibitor tablet (Roche) to 10ml buffer.)

    For Antibody ab36977 and ab78054 I also used a homogenate based on the RIPA buffer according to your suggestions. As you can see below in the previous mail, there is basically no difference between the two homogenates, why I only used the Tris-buffer based for this testing as most of our homogenates are based on that buffer.



    My problems with the ab17044 is that it gives several bands all very high in the gel. You have previously written to me:

    “Regarding the multiple bands with ab36977. This protein undergoes modifications such as glycosylation and palmitoylation which can certainly shift the bands you observe to ˜75 - 80kDa. The ˜50kDa band is likely unmodified protein”

    Which make sense, but now the bands are situated even higher in the gel:

    I am unsure whether the very intense band above 100kDa are specific?? If I block the signal (by laying something over the membrane when the picture is taken) I get this result, which actually is quite nice, but have I by that removed some CD36 signal? The middle part of the membrane was used for testing another antibody without success.

    CD 36 on Lane A-D: all with 15µg total protein.

    Lane A:

    Blocking for 1½ hour in 5% BSA in TBST at RT

    overnight incubation at 4 degrees with antibody diluted in 5% BSA in TBST with agitation (in a tube rolling) with a 1:500 dilution of ab17044.



    Lane B:

    Blocking for 1½ hour in 5% BSA in TBST at RT

    overnight incubation at 4 degrees with antibody diluted in 5% BSA in TBST with agitation (in a tube rolling) with a 1:1000 dilution of ab17044.



    Lane C:

    Blocking for 1½ hour in 3% BSA in TBST at RT

    overnight incubation at 4 degrees with antibody diluted in 3% BSA in TBST with agitation (in a tube rolling) with a 1:500 dilution of ab17044.



    Lane D:

    Blocking for 1½ hour in 3% BSA in TBST at RT

    overnight incubation at 4 degrees with antibody diluted in 3% BSA in TBST with agitation (in a tube rolling) with a 1:1000 dilution of ab17044.



    All lanes

    Sec. antibody was diluted in 5% BSA in TBST and incubated for 2 hours at 25 degrees in a tube rolling.



    All wash steps was in TBST. Both membranes was incubated with ECL and picture was taken with a CCD camera in 1 min.



    How many of the bands are specific antibody reactions with CD36 proteins??



    Kind regards

    Read More
    Answer

    Thank you for taking the time to send us your results. I am sorry to hear you have had difficulty obtaining satisfactory results from this CD36 antibody as well.

    The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

    The bands you observe above 100 kDa are likely to be unspecific. Reviewing this case, it seems your sample has not migrated sufficiently into the gel which would also explain that your testing for another antibody failed.

    I would like to offer some suggestions to help optimise the results you obtained from ab17044. I would also appreciate if you can confirm some further details:

    1) Your target is amembrane protein and it is heavily modified (as discussed before), and these proteins have the unfortunate tendency to aggregate during boiling periods with high temperatures and beta- mercaptoethanol (bME) due to their hydrophobic nature. I would like to suggest to use DTT as reducing agent rather than bME and to decrease the boiling temperature to 65ºC and to prolong the boiling time up to 10 min.

    2) What gel percentage have you used and which buffer system? For this target 8-10% gels might be sufficient. Also, the buffer system can have an influence thesize at which a bandruns (see attached).

    3) To verify the transfer of the proteins to the membrane I would recommend the Ponceau staining, if you have not already tried (see protocol details attached).

    I would hope that these suggestions help to optimise the results and would lead to distinct bands for CD36. It would then not be necessary to 'block out' the unspecific signals (which is also not really common scientific practise). Usually, the specific signal of an antibody is blocked by incubating the antibodywith the immunogenic peptide prior to staining the membrane. This way the specific signal disappears whereas unspecific background staining would still be present, validating the results. Unfortunately, the exact epitope of this particular CD36 antibody is unknown, and therefore, this blocking of the signal is not an option.

    I hope this information is helpful for you though, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

    Read More
    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Blocking
    Sample
    Mouse Cell (mouse macrophage cell line(raw264.7 cell))
    Specification
    mouse macrophage cell line(raw264.7 cell)

    Abcam user community

    Verified customer

    Submitted Nov 27 2014

    Answer

    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this our Anti-CD36 antibody [FA6-152] (ab17044) .

    The details you have kindly provided will provide us with vital information for our monitoring of product quality

    I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there are no further tips to provide on this occasion to help improve the results. I can confirm that the vial might not be working for you because of the circumstances.

    We discovered from our source that this antibody is stable at -20 degrees. Therefore we now recommend to store it at this temperature.

    I apologise for the inconvenience and am pleased to offer you a free of charge replacement of the same product, a credit note or refund in compensation.

    In addition, I can suggest you may like to consider the following suggestions as a check for future experiments:

    We recommend a positive control to confirm the quality of the primary antibody. For the Anti-CD36 antibody [FA6-152] (ab17044) we can recommend HEL cells.

    Furthermore we suggest to make sure that the secondary antibody is raised against the isotype of the primary antibody if not already done. Additionally it would be useful to confirm that the secondary antibody is working well with other primary antibodies.

    Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Answer

    Thank you for contacting us.

    The immunogen for ab17044 is 20 weeks old fetal erythrocytes. However, we have not further mapped the epitope for this antibody.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    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 a free of charge replacement with the order number xxx with ab64014.

    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
    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 a free of charge replacement with the order number 1135221.

    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
    Answer

    Thank you for filling in the questionnaire. I am sorry to hear that you are still experiencing problems with this antibody.
    I have looked through your protocol and would like to ask you some questions in order to understand your procedure better as well as make a few suggestions to improve your results:
    - Could you please provide further details about the sample type used? What kind of sample homogenate was used, whole cell lysate, purified protein?
    - This particular protein shows different sites of glycosilation, producing bands at higher molecular weights, and so, sample preparation is crucial. Make sure the protein is running under denatured / reducing conditions.
    - Have you checked with other techniques the negative control does not contain this cell marker?
    - Have you run a non primary control to dismiss the non specific band corresponds to the secondary?
    It would be really helpful if you could provide us with some images from any of the western blots performed, to try to find out why the antibody did not perform as stated on the datasheet.
    Could you also please provide the order number? As our Abpromise states, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.
    I look forward to receiving your reply. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question
    Answer

    Thank you for contacting us.

    According to our information, we have that platelet aggregation and secretion was induced by > 1μg/ml of this antibody, according to this reference: Kieffer N et al; Development regulated expression of a 78 kDa erythroblast membrane glycoprotein immunologically related to the platelet thrombospondin receptor. Biochem J 1989, 262: 835

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Answer

    Thank you for contacting us.
    I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.
    In order to better understand the problem, I would like to send you a questionnaire with some questions about the protocol used, as well as the order details. All this information is crucial to help us understand the possible causes of the problem. Any images are highly appreciated.
    Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. If no suggestions can be made to the protocol, and the antibody was purchased within the last 6 months, it is covered by our Abpromise guarantee, and therefore I can offer you a free of charge replacement or a credit note for it.
    I look forward to receiving your reply. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Answer

    Thank you for contacting us. Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

    We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

    If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. Please contact our Scientific Support team by replying to this email prior to purchase for more information.

    Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

    To find out more about our Abreview system, please see the following link:

    https://www.abcam.com/abreviews

    I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

    Read More

    1-10 of 23 Abreviews or Q&A

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