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Key Product Details

Validated by

Orthogonal Validation, Biological Validation

Species Reactivity

Human, Mouse, Rat, Porcine, Bovine, Primate, Rabbit

Applications

Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free

Concentration

1 mg/ml

Product Summary for CD36 Antibody - BSA Free

Immunogen

This CD36 Antibody was developed against a synthetic peptide mapping to a region of human CD36 between residues 100-200 [Uniprot# P16671]

Reactivity Notes

Use in Mouse reported in scientific literature (PMID:34974159) Porcine reactivity reported in scientific literature (PMID: 23727393). Rat reactivity reported in scientific literature (PMID: 25635851). Rabbit reactivity reported in scientific literature (PMID: 30105261).

Localization

Integral membrane

Marker

Endothelial Cell Marker

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

110 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.

Scientific Data Images for CD36 Antibody - BSA Free

Immunohistochemistry: CD36 Antibody [NB400-144] - Timp4-deficiency results in defective lipid digestion and absorption. (B) Immunostaining for CD36 in small intestine (proximal region) of chow-fed and HFD-fed WT and Timp4-/- mice. Western blot (C) and mRNA (D) for CD36 in enterocyte fraction of chow-fed and HFD-fed WT and Timp4-/- mice (collected from the proximal small intestine). Image collected and cropped by CiteAb from the following publication (http://www.nature.com/articles/s41598-017-05951-4), licensed under a CC-BY license.
Western Blot: CD36 Antibody [NB400-144] - Total protein from Human Skin and Adipose tissue, Mouse Adipose and Rat Adipose tissue was separated on a 12% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 2.0 ug/ml anti-CD36 in 5% non-fat milk in TBST and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.
Immunohistochemistry-Paraffin: CD36 Antibody [NB400-144] - Staining of CD36 in human adipocytes.

Applications for CD36 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:50-1:200

Immunohistochemistry

1:200-1:400

Immunohistochemistry-Frozen

reported in scientific literature (PMID 24531551)

Immunohistochemistry-Paraffin

1:200-1:400

Western Blot

1:500-1:2000
Application Notes
In Western Blot, a band is seen ~75-80 kDa. The theoretical molecular weight of CD36 is ~53 kDa. The difference in theoretical MW and actual MW as seen in Western blot is most likely due to the heavy glycosylation and palmitoylation of this protein. 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.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 10 reviews rated 3.8 using NB400-144 in the following applications:

Published Applications

Read 124 publications using NB400-144 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: CD36

Originally discovered in platelets, cluster of differentiation 36, CD36, (also known as thrombospondin receptor, fatty acid translocase (FAT), platelet membrane glycoprotein IV (GPIV), and scavenger receptor class B, member 3 (SR-B3)) is a plasma membrane glycoprotein belonging to the class B scavenger receptor family (1,2). Human, mouse, and rat CD36 is synthesized as a 472 amino acid (a.a.) protein with a theoretical molecular weight of 53 kDa for the canonical isoform (3). Its domains include a short cytoplasmic tail at the N-terminal and C-terminal and a large extracellular loop flanked on each side by a transmembrane domain. The extracellular domain facilitates the update of fatty acids (FFAs), phospholipids, and cholesterol by forming two hydrophobic cavities, which was first modeled in the CD36 homologue, LIMP-2/ SCARB2 (4).

The expression of CD36 has been reported in platelets, erythrocytes, monocytes, differentiated adipocytes, skeletal muscle, mammary epithelial cells, spleen cells, some skin microdermal endothelial cells, and in cancer. Circulating levels of soluble CD36 (sCD36) has also been reported in chronic inflammatory disease such as type 2 diabetes and chronic kidney disease. CD36 participates in angiogenesis, innate immunity, and the clearance of apoptotic phagocytes. In lipid metabolism, CD36 functions as a macrophage receptor for oxidized LDL and as an adipocyte receptor/transporter for long-chain FFAs. Plasmodium falciparum, the parasite that causes malaria, binds CD36 via PfEMP1 proteins, tethering parasite-infected erythrocytes to endothelial receptors (5). Anti-CD36 isoantibodies have been detected in Type 1 CD36-deficient mothers and is implicated as the cause of fetal/neonatal alloimmune thrombocytopenia (6).

References

1) Febbraio, M., Hajjar, D. P., & Silverstein, R. L. (2001). CD36: a class B scavenger receptor involved in angiogenesis, atherosclerosis, inflammation, and lipid metabolism. The Journal of clinical investigation, 108(6), 785-791. PMID: 11560944

2) Silverstein RL, Febbraio M. (2000) CD36 and atherosclerosis. Curr Opin Lipidol. 2000 11(5):483-91. PMID: 11048891.

3) Endemann G, Stanton LW, Madden KS, Bryant CM, White RT, Protter AA. (1993) CD36 is a receptor for oxidized low density lipoprotein. J Biol Chem. 268(16):11811-6. PMID: 7685021.

4) Wang, J., & Li, Y. (2019). CD36 tango in cancer: signaling pathways and functions. Theranostics, 9(17), 4893-4908. PMID: 31410189

5) Hsieh FL, Turner L, Bolla JR, Robinson CV, Lavstsen T, Higgins MK. (2016) The structural basis for CD36 binding by the malaria parasite. Nat Commun. 7:12837. PMID: 27667267

6) Gruarin P, Ulliers D, Thorne RF, Alessio M. (2000) Methionine 156 in the immunodominant domain of CD36 contributes to define the epitope recognized by the NL07 MoAb. Mol Cell Biochem 214, 115-121. PMID: 11195795.

Alternate Names

BDPLT10, CD36 antigen (collagen type I receptor, thrombospondin receptor), CD36 molecule (thrombospondin receptor), CHDS7, cluster determinant 36, FAT, Fatty acid translocase, Glycoprotein IIIb, GP3B, GP4, GPIIIB, GPIV, leukocyte differentiation antigen CD36, PAS IV, PAS-4 protein, PASIV, platelet glycoprotein 4, platelet glycoprotein IV, SCARB3, scavenger receptor class B, member 3

Gene Symbol

CD36

Product Documents for CD36 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for CD36 Antibody - BSA Free

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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Protocols

View specific protocols for CD36 Antibody - BSA Free (NB400-144):

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and wash the cells briefly in PBS. Add 10% formalin to the dish and fix at room temperature for 10 minutes.
2. Remove the formalin and wash the cells in PBS.
3. Permeablize the cells with 0.1% Triton X100 or other suitable detergent for 10 min.
4. Remove the permeablization buffer and wash three times for 10 minutes each in PBS. Be sure to not let the specimen dry out.
5. To block nonspecific antibody binding, incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
6. Add primary antibody at appropriate dilution and incubate overnight at 4C.
7. Remove primary antibody and replace with PBS. Wash three times for 10 minutes each.
8. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
9. Remove secondary antibody and replace with PBS. Wash three times for 10 minutes each.
10. Counter stain DNA with DAPi if required.
Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.

Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions.

FAQs for CD36 Antibody - BSA Free

Showing  1 - 2 of 2 FAQs Showing All
  • Q: Can you tell me if this product, NB400-144, requires antigen retrieval for hick on paraffin embedded tissues? If so, what type of antigen retrieval is recommended?

    A: Yes, we recommend antigen retrieval for IHC-P with NB400-144 and in our testing we performed heat-mediated antigen retrieval with sodium citrate buffer (pH 6).

  • Q: We used the NB400-144 to detect CD36 in guinea pig by Western blot analysis. Is there a similar antibody recommended for IP?

    A: Here is a link to other CD36 antibodies which are validated for IP. The CD36 antibody (NB400-144) that you already have is one of our best selling products with over 15 PubMed citations. We have not tested this antibody in IP yet and I do not see any reason why it would not work in IP. If you would be interested in testing this novel application, please take a look at our Innovator's Reward program.

Showing  1 - 2 of 2 FAQs Showing All
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