Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Immunocytochemistry

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 706824
View all N-Acetylglucosaminyltransferase V/MGAT5 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human N-Acetylglucosaminyltransferase V/MGAT5
Leu189-Leu741
Accession # Q09328

Specificity

Detects human N-Acetylglucosaminyltransferase V/MGAT5 in direct ELISAs and Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Human N‑Acetylglucosaminyltransferase V/MGAT5 Antibody

Detection of Human N-Acetylglucosaminyltransferase V/MGAT5 antibody by Western Blot.

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 antibody in MCF-7 Human Cell Line by Immunocytochemistry (ICC).

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.
Detection of N-Acetylglucosaminyltransferase V/MGAT5 by Western Blot

Detection of N-Acetylglucosaminyltransferase V/MGAT5 by Western Blot

Knockdown of Mgat5 expression in cultured human RPE cells attenuates binding of Gal-3 to RPE cells.(A) Western blot analysis of Mgat5 expression in RPE cells transfected with siRNA containing the same nucleotides as Mgat5 siRNA in random order (sc siRNA), and the same cell line transfected with 50 pmol and 100 pmol of double stranded siRNA complementary to Mgat5 (Mgat5 siRNA), respectively. Lysates containing approximately equal amounts of protein were separated by SDS-PAGE and blotted for immunochemical detection of Mgat5 content. Experiments were repeated at least three times. MW; molecular weight. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26760037), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of N-Acetylglucosaminyltransferase V/MGAT5 by Western Blot

Detection of N-Acetylglucosaminyltransferase V/MGAT5 by Western Blot

Knockdown of Mgat5 expression in cultured human RPE cells attenuates binding of Gal-3 to RPE cells. The target sequence derived from the genomic sequence of Mgat5 was inserted into a CRISPR-Cas9 nuclease expressing lentiviral vector and ARPE19 cells were transfected using Lipfectamine 2000 Plus reagent. (D) Western blot analysis of Mgat5 expression in ARPE19 cells transduced with guide RNA leading to specific knockdown of the Mgat5 (gMgat5), or cells tranduced with a CRISPR-Cas9 lentiviral vector encoding for an none-coding filler RNA (LV), or wild-type ARPE-19 cells. Lysates containing approximately equal amounts of protein were separated by SDS-PAGE and blotted for immunochemical detection of Mgat5 content. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26760037), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human N-Acetylglucosaminyltransferase V/MGAT5 by Immunohistochemistry

Detection of Human N-Acetylglucosaminyltransferase V/MGAT5 by Immunohistochemistry

Clinical significance of MGAT5 as a potential therapeutic target in tumour EMT. A Representative IHC staining of MGAT5 in CRC patient tissue from the TMA (ZL-Cocsur1241). B MGAT5 protein expression in malignant colorectal cancer (T) and adjacent nonmalignant tissues (N). The data are presented as a box plot. Whiskers show the min-to-max values, n = 69 per group. C Kaplan‒Meier analysis of OS in patients with varying MGAT5 expression levels based on data from selected CRC tissues. D Kaplan‒Meier analyses were performed to assess the associations between MGAT5 expression and RFS. E MGAT5 protein levels in nonmetastatic and metastatic colorectal cancer (T) tissues. The data are presented as a box plot. The whiskers show the min-to-max values. F Immunostaining of the CRC tissue TMA for MGAT5 or ZO-1. Correlations between the mean MGAT5 intensity and the mean ZO-1 intensity in CRC tissue samples (n = 15). Pearson's correlation analysis yielded the R and p values. Scale bar: 100 µm Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39511539), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human N‑Acetylglucosaminyltransferase V/MGAT5 Antibody

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed MCF‑7 human breast cancer cell line

Western Blot

2 µg/mL
Sample: HepG2 human hepatocellular carcinoma cell line

Reviewed Applications

Read 1 review rated 4 using MAB5469 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: N-Acetylglucosaminyltransferase V/MGAT5

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).

References

  1. Saito, H. et al. (1994) Biochem. Biophys. Res. Commun. 198:318.
  2. Lau, K.S. et al. (2007) Cell 198:123.
  3. Dennis, J.W. et al. (2002) Biochim. Biophys. Acta 1573:414.
  4. Granovsky, M. et al. (2008) Nat. Med. 7:1.
  5. Kim, Y.S. et al. (2008) Mol. Cell. Proteomics 7:1.

Long Name

alpha-1,6-Mannosylglycoprotein 6-beta-N-acetylglucosaminyltransferase A

Alternate Names

MGAT5, N-Acetylglucosaminyltransferase V

Entrez Gene IDs

4249 (Human); 107895 (Mouse); 65271 (Rat)

Gene Symbol

MGAT5

UniProt

Q09328 (Human)

Additional N-Acetylglucosaminyltransferase V/MGAT5 Products

Product Documents for Human N‑Acetylglucosaminyltransferase V/MGAT5 Antibody

Certificate of Analysis

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Product Specific Notices for Human N‑Acetylglucosaminyltransferase V/MGAT5 Antibody

For research use only

Related Research Areas

Glycobiology-related Enzymes

Citations for Human N‑Acetylglucosaminyltransferase V/MGAT5 Antibody

Customer Reviews for Human N‑Acetylglucosaminyltransferase V/MGAT5 Antibody (1)

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  • Human N-Acetylglucosaminyltransferase V/MGAT5 Antibody
    Name: SOK LEE
    Application: Western Blot
    Sample Tested: Cell Lysates
    Species: Human
    Verified Customer | Posted 03/23/2018
    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 4C Secondary Ab: anti-mouse HRP antibody (1:10000 dilution) 90 minute at room temperature. Development: SuperSignal West-Pico PLUS for 15 sec.
    Human N‑Acetylglucosaminyltransferase V/MGAT5 Antibody MAB5469

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