Detection of Human N‑Acetylglucosaminyltransferase V/MGAT5 by Western Blot.
Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human N‑Acetylglucosaminyltransferase V/MGAT5 Monoclonal Antibody (Catalog # MAB5469) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for N‑Acetylglucosaminyltransferase V/MGAT5 at approximately 100 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
N‑Acetylglucosaminyltransferase V/MGAT5 in MCF‑7 Human Cell Line.
N‑Acetylglucosaminyltransferase V/MGAT5 was detected in immersion fixed MCF‑7 human breast cancer cell line using Mouse Anti-Human N‑Acetylglucosaminyltransferase V/MGAT5 Monoclonal Antibody (Catalog # MAB5469) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to plasma membranes and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
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.
N-Acetylglucosaminyltransferase V (GnT-V), also known as mannosylglycoprotein N-acetyl-glucosaminyltransferase 5 (MGAT5), adds an N-acetylglucosamine to the alpha 1-6-linked core mannose of an N-linked oligosaccharide in the Golgi apparatus (1). This reaction is the committing step for the biosynthesis of beta 1-6GlcNAc-branched arm in N-glycans. The degree of N-glycan branching has been shown to regulate cell proliferation and differentiation (2). An increase in the GnT-V activity and its glycan products is also known to positively correlate with the progression of invasive malignancies (3, 4). For example, ectopic expression of GnT-V in epithelial cells results in morphological transformation and tumor growth in mice and overexpression in carcinoma cells has been shown to induce metastatic spread (3-5).
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Lau, K.S. et al. (2007) Cell 198:123.
Dennis, J.W. et al. (2002) Biochim. Biophys. Acta 1573:414.
Granovsky, M. et al. (2008) Nat. Med. 7:1.
Kim, Y.S. et al. (2008) Mol. Cell. Proteomics 7:1.
alpha-1,6-Mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A
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Western Blot: Human N-Acetylglucosaminyltransferase V/MGAT5 Antibody [MAB5469].
Other Experimental Details
Other Experimental Details
MGAT5 western blotting after MGAT5A transfection to 293 cells(Figure Legend)Ctrl: Control vector was transfected for 48 hrs.MGAT5: MGAT5A vector was transfected for 48 hrs.Above blots: human MGAT5 (80-100 kDa)Lower blots: beta-actin (42-kDa)Western blot condition: Tris buffer (0.5% Triton X)-lysed cell extract (15 ug each) was loaded with non-reducing condition (:No beta-MercaptoEthanol on loading buffer).Primary Ab: anti-MGAT5 antibody (1 ug/ml) overnight at 4CSecondary Ab: anti-mouse HRP antibody (1:10000 dilution) 90 minute at room temperature.Development: SuperSignal West-Pico PLUS for 15 sec.