Detection of Human Galectin‑3BP/MAC‑2BP by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Human Galectin‑3BP/MAC‑2BP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2226) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Galectin‑3BP/MAC‑2BP at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Galectin‑3BP/MAC‑2BP in HeLa Human Cell Line.
Galectin‑3BP/MAC‑2BP was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human Galectin‑3BP/MAC‑2BP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2226) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human Galectin‑3BP/MAC‑2BP by Simple WesternTM.
Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and HepG2 human hepatocellular carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Galectin‑3BP/MAC‑2BP at approximately 115 kDa (as indicated) using 10 µg/mL of Goat Anti-Human Galectin‑3BP/MAC‑2BP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2226) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.
Galectin-3 binding protein (Galectin-3BP), also known as MAC-2 binding protein (MAC-2BP or M2BP), and the 90 kDa tumor associated antigen (TAA90K or 90K), is a secreted glycoprotein of the scavenger receptor cysteine-rich (SRCR) superfamily (1, 2). Galectin-3BP binds Galectin-3 (formerly MAC-2) with high affinity, but also binds Galectins -1 and -7, several collagen types, fibronectin, beta 1 integrins and nidogen (3, 6, 7). It is widely expressed in all extracellular fluids and in pericellular areas of cell-rich tissues (1-3). The 585 amino acid (aa) human Galectin-3BP contains an 18 aa signal sequence and four definitive domains (4-6). Domain 1 is a group A scavenger receptor domain (4), domain 2 is a BTB/POZ domain that strongly mediates dimerization (5), and domain 3 is an IVR domain, that is also found following the POZ domain in Drosophila kelch protein. Although little is known about domain 4, recombinant domains 3 and 4 reproduce the solid-phase adhesion profile of full-length Galectin-3BP (5, 6). Glycosylation at seven N-linked sites, generates a molecular size of 85-97 kDa (1, 2, 6). Galectin-3BP dimers form linear and ring-shaped oligomers, most commonly decamers and dodecamers (3, 5). In vitro, Galectin-3BP has been shown to stimulate natural killer cells and lymphokine-activated killer cell activity (2). High Galectin-3BP expression has been correlated with tumor aggressiveness in several, but not all, study systems (7). Mature human Galectin-3BP shares 69% aa identity with mouse cyclophilin C-associated protein (CyCAP), which does not appear to bind Galectin-3 (8). Human Galectin-3BP also shares 73%, 67% and 68% aa identity with relatively uncharacterized orthologs in dog, rat and cow, respectively. A human N-terminally truncated sequence that begins within the BTB/POZ domain (aa 196) has been reported.
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Jalkanen, K. et al. (2001) Eur. J. Immunol. 31:3075.
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Extracellular matrix proteomics identifies molecular signature of symptomatic carotid plaques
Authors: SR Langley, K Willeit, A Didangelos, LP Matic, P Skroblin, J Barallobre, M Lengquist, G Rungger, A Kapustin, L Kedenko, C Molenaar, R Lu, T Barwari, G Suna, X Yin, B Iglseder, B Paulweber, P Willeit, J Shalhoub, G Pasterkamp, AH Davies, C Monaco, U Hedin, CM Shanahan, J Willeit, S Kiechl, M Mayr
J. Clin. Invest, 2017;0(0):.
Sample Type: Whole Tissue
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