EMMPRIN/CD147 was detected in immersion fixed paraffin-embedded sections of human colon using Mouse Anti-Human EMMPRIN/CD147 Monoclonal Antibody (Catalog # MAB9721) at 1 µg/mL at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 647 Goat anti-Mouse IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane. Protocol available in COMET™ Panel Builder.

Detection of EMMPRIN/CD147 in Human Liver Cancer via seqIF™ staining on COMET™

EMMPRIN/CD147 was detected in immersion fixed paraffin-embedded sections of human liver cancer using Mouse Anti-Human EMMPRIN/CD147 Monoclonal Antibody (Catalog # MAB9721) at 1 µg/mL at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 647 Goat anti-Mouse IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane. Protocol available in COMET™ Panel Builder.

Detection of EMMPRIN/CD147 in Human Pancreatic Cancer via seqIF™ staining on COMET™

EMMPRIN/CD147 was detected in immersion fixed paraffin-embedded sections of human pancreatic cancer using Mouse Anti-Human EMMPRIN/CD147 Monoclonal Antibody (Catalog # MAB9721) at 1 µg/mL at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 647 Goat anti-Mouse IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane. Protocol available in COMET™ Panel Builder.

Detection of Human EMMPRIN/CD147 by Western Blot.

Western blot shows lysates of human brain (cerebellum), human heart (ventricle), and human liver. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human EMMPRIN/CD147 Monoclonal Antibody (Catalog # MAB9721) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for EMMPRIN/CD147 at approximately 45-60 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

EMMPRIN/CD147 in 789-O Human Cell Line.

EMMPRIN/CD147 was detected in immersion fixed 789-O Human renal cell carcinoma cell line using Mouse Anti-Human EMMPRIN/CD147 Monoclonal Antibody (Catalog # MAB9721) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cell membrane. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

EMMPRIN/CD147 in Human Heart.

EMMPRIN/CD147 was detected in immersion fixed paraffin-embedded sections of human heart using Mouse Anti-Human EMMPRIN/CD147 Monoclonal Antibody (Catalog # MAB9721) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell membrane in capillaries. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Western Blot Shows Human EMMPRIN/CD147 Specificity by Using Knockout Cell Line.

Western blot shows lysates of HEK293T human embryonic kidney cell line and human EMMPRIN knockout HEK293T human embryonic kidney cell line (KO). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human EMMPRIN/CD147 Monoclonal Antibody (Catalog # MAB9721) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for EMMPRIN/CD147 at approximately 45-60 kDa (as indicated) in the parental cell line, but is not detectable in knockout cell line. GAPDH (MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

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Applications for Human EMMPRIN/CD147 Antibody

Application

Recommended Usage

COMET

1 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon, liver cancer and pancreatic cancer.

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed 789-O Human renal cell carcinoma cell line

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human heart

Knockout Validated

2 µg/mL
Sample: EMMPRIN/CD147 is specifically detected in HEK293T human embryonic kidney parental cell line but is not detectable in EMMPRIN/CD147 knockout HEK293T human embryonic kidney cell line.

Western Blot

2 µg/mL
Sample: Human brain (cerebellum), human heart (ventricle), and human liver

Reviewed Applications

Read 1 review rated 5 using MAB9721 in the following applications:

Immunohistochemistry (1 Review)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: EMMPRIN/CD147

Extracellular matrix metalloproteinase (MMP) inducer (EMMPRIN), also known as basigin and CD147, is a 44-66 kDa, variably N- and O-glycosylated, type I transmembrane protein that belongs to the immunoglobulin superfamily (1-4). Human EMMPRIN is 269 amino acids (aa) in length and contains a 24 aa signal sequence, a 183 aa extracellular domain (ECD), a 21 aa transmembrane (TM) segment and a 41 aa cytoplasmic tail. The ECD contains one C2-type and one V-type Ig-like domain. There is one 385 aa splice variant that contains an extra N-terminal IgCAM domain and is found only in the retina (5). There are additional multiple potential isoform variants for EMMMPRIN. Two that have been characterized are 205 and 176 aa in length. The 176 aa isoform utilizes an alternative start site at Met94, while the 205 aa isoform contains an 11 aa substitution for aa 1-75. Notably, the 176 aa isoform heterodimerizes with the standard EMMPRIN isoform and down-modulates its activity. This is in contrast to EMMPRIN homodimers that show full biological activity (6). EMMPRIN is expressed in areas of tissue remodeling, including endometrium, placenta, skin, and regions undergoing angiogenesis (1, 2, 7-10). It is also expressed on cells with high metabolic activity, such as lymphoblasts, macrophages and particularly tumor cells (2, 11). On such cells, EMMPRIN is often co-expressed with the amino acid transporter CD98h (12). EMMPRIN also interacts with caveolin-1 (via its C2-like domain), and this reduces the level of EMMPRIN glycosylation and subsequent EMMPRIN multimerization and activity (13). In addition, EMMMPRIN is reported to complex with both annexin II and beta1 integrins alpha3 and alpha6, an interaction that contributes to tumor growth and metastasis (14-16). Finally, the soluble calcium-binding protein S100A9 has now been identified as a ligand for EMMPRIN, and may mediate many of the tumorigenic activities attributed to EMMPRIN (17). EMMPRIN's TM sequence contains a charged aa (Glu), and a Pro important for intracellular interactions with cyclophilins (CyP) (3, 18, 19). CyPA (cyclosporin A receptor) and CyP60 interactions with the TM segment promote leukocyte inflammatory chemotaxis and surface expression of EMMPRIN, respectively (18, 19). An active 22 kDa fragment can be shed from tumor cells by MT1-MMP (1). Tumor cells can also release active, full-length EMMPRIN in microvesicles (20, 21). Functionally, EMMPRIN is known to induce urokinase-type plasminogen activator (uPA), VEGF, hyaluronan and multiple MMPs (1, 2, 8-10). Human EMMPRIN (269 aa) shows 58%, 58%, 62% and 52% aa identity with mouse, rat, cow and chick EMMPRIN, respectively. It also shows 25% and 38% aa identity with the related proteins, embigin and neuroplastin (SDR-1), respectively. SARS-CoV-2 invades host cells via two receptors: angiotensin-converting enzyme 2 (ACE2) and EMMPRIN. Spike protein (SP) from virus binds to ACE2 or EMMPRIN on the host cell, mediating viral invasion and dissemination of virus among other cells (22). EMMPRIN is a second entry receptor for SARS-CoV-2 (22). It is present in multiple cellular types in lung and highly expressed in type II pneumocytes and macrophages at the edges of the fibrotic zones (22). Therefore, the blockade of EMMPRIN could also play a beneficial role in pulmonary fibrosis due to COVID-19 (22).

References

Gabison, E. E. et al. (2005) Biochimie 87:361.
Yurchenko, V. et al. (2006) Immunology 117:301.
Kasinrerk, W. et al. (1992) J. Immunol. 149:847.
Iacono, K.T. et al. (2007) Exp. Mol. Pathol. 83:283.
Hanna, S. M. et al. (2003) BMC Biochem. 4:17.
Liao, C-G. et al. (2011) Mol. Cell. Biol. 31:2591.
Riethdorf, S. et al. (2006) Int. J. Cancer 119:1800.
Braundmeier, A. G. et al. (2006) J. Clin. Endocrinol. Metab. 91:2358.
Tang, Y. et al. (2006) Mol. Cancer Res. 4:371.
Quemener, C. et al. (2007) Cancer Res. 67:9.
Wilson, M. C. et al. (2005) J. Biol. Chem. 280:27213.
Xu, D. and M. E. Hemler, (2005) Mol. Cell. Proteomics 4:1061.
Tang, W. et al. (2004) Mol. Biol. Cell 15:4043.
Zhao, P. et al. (2010) Cancer Sci. 101:387.
Dai, J. et al. (2009) BMC Cancer 9:337.
Li, Y. et al. (2012) J. Biol. Chem. 287:4759.
Hibino, T. et al. (2013) Cancer Res. 73:172.
Arora, K. et al. (2005) J. Immunol. 175:517.
Pushkarsky, T. et al. (2005) J. Biol. Chem. 280:27866.
Egawa, N. et al. (2006) J. Biol. Chem. 281:37576.
Sidhu, S. S. et al. (2004) Oncogene 23:956.
Ulrich, H. et al. (2020) Stem Cell Rev. and Rep., https://doi.org/10.1007/s12015-020-09976-7.

Long Name

Extracellular Matrix Metalloproteinase Inducer

Alternate Names

Basigin, BSG, CD147

Entrez Gene IDs

682 (Human); 12215 (Mouse); 102121721 (Cynomolgus Monkey)

Gene Symbol

BSG

UniProt

P35613.2

Additional EMMPRIN/CD147 Products

Product Documents for Human EMMPRIN/CD147 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human EMMPRIN/CD147 Antibody

For research use only

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