O-GlcNAc Antibody (RL2) - BSA Free

Immunohistochemistry-Paraffin: O-GlcNAc Antibody (RL2) [NB300-524] - Analysis of a FFPE tissue section of the mouse colon using 1:200 dilution of O-GlcNAc [RL2] antibody (NB300-524). The signal was developed using HRP-DAB method which followed counterstaining of the cells with hematoxylin.

Western Blot: O-GlcNAc Antibody (RL2) [NB300-524] - Impact of OGA inhibition by thiamet G in primary cultures of NCM. Representative western blots (left panel) and quantification (right panel) of O-GlcNAcylated proteins levels in control (Ctrl) and NCM treated with 100 nM of thiamet G (TG) during 24 h (n = 12). Image collected and cropped by Citeab from the following publication (Interplay Between Phosphorylation and O-GlcNAcylation of Sarcomeric Proteins in Ischemic Heart Failure. Front Endocrinol (Lausanne) (2018) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: O-GlcNAc Antibody (RL2) [NB300-524] - Neuro2a cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton X-100. The cells were incubated with anti-O-GlcNAc (RL2) at 5 ug/mL overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Flow Cytometry: O-GlcNAc Antibody (RL2) [NB300-524] - An intracellular stain was performed on Neuro2a cells with O-GlcNAc Antibody [RL2] NB300-524 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Western Blot: O-GlcNAc Antibody (RL2) [NB300-524] - Analysis of mouse cortical brain lysates using O-Linked N-Acetylglucosamine Monoclonal Antibody. Blots containing cortical extracts from 4 individual C57BL/6 mice (Lanes 1-4) were blocked with 5% milk in TBST, and probed with MA1-072 at 1:1000, followed by a fluorophore-conjugated goat anti-mouse IgG secondary antibody. Data courtesy of the Innovators Program.

Immunocytochemistry/Immunofluorescence: O-GlcNAc Antibody (RL2) [NB300-524] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.5% Triton X-100. The cells were incubated with anti-O-GlcNAc (RL2) at 5 ug/mL overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Flow Cytometry: O-GlcNAc Antibody (RL2) [NB300-524] - Analysis using Alexa Fluor (R) 647 conjugate of NB300-524. An intracellular stain was performed on Jurkat cells with O-GlcNAc antibody (RL2) NB300-524 (blue) and a matched isotype control NBP2-27287 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. 1 ug of antibody was added to 100 uL of staining buffer and cells were incubated for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Flow Cytometry: O-GlcNAc Antibody (RL2) [NB300-524] - An intracellular stain was performed on U-937 cells with O-GlcNAc antibody (RL2) NB300-524AF647 (blue) and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature . Both antibodies were conjugated to Alexa Fluor 647.

Flow Cytometry: O-GlcNAc Antibody (RL2) [NB300-524] - An intracellular stain was performed on Jurkat cells with O-GlcNAc antibody (RL2) NB300-524PE (blue) and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature . Both antibodies were conjugated to Phycoerythrin.

Flow Cytometry: O-GlcNAc Antibody (RL2) [NB300-524] - An intracellular stain was performed on SK-MEL-28 cells with O-GlcNAc antibody (RL2) NB300-524AF647 (blue) and a matched isotype control. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature . Both antibodies were conjugated to Alexa Fluor 647.

Flow Cytometry: O-GlcNAc Antibody (RL2) [NB300-524] - An intracellular stain was performed on HeLa cells with O-GlcNAc Antibody [RL2] Antibody NB300-524AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Flow Cytometry: O-GlcNAc Antibody (RL2) [NB300-524] - An intracellular stain was performed on Neuro2a cells with O-GlcNAc Antibody [RL2] NB300-524AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Flow Cytometry: O-GlcNAc Antibody (RL2) [NB300-524] - An intracellular stain was performed on RH30 cells with O-GlcNAc [RL2] Antibody NB300-524AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Flow Cytometry: O-GlcNAc Antibody (RL2) [NB300-524] - An intracellular stain was performed on Jurkat cells with O-GlcNAc Antibody [RL2] NB300-524 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 1.0 ug/mL for 30 minutes at room temperature, followed by Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (35503, Thermo Fisher).

Western Blot: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524] - Analysis of O-GlcNAcylated LV proteins by Western blot & WGA-SDS-PAGE gel electrophoresis. (A) Red ponceau staining (left panel) & western blot (right panel) of O-GlcNAcylated proteins (50 μg) extracted from sham- & HF-rats treated or not with thiamet G. The positions of molecular weight are indicated as kilodalton (kDa) on the left. (B) Red ponceau staining (left panel) & WGA-SDS-PAGE of O-GlcNAcylated proteins levels (middle panel) of O-GlcNAcylated desmin levels (right panel) from the same samples. The arrow in desmin WGA gels indicates the non-O-GlcNAcylated form. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30344511), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524] - Effect of OGA inhibition by thiamet G in isolated perfused heart. (A) Description of the protocol designed for thiamet G (TG) perfusion in sham- (n = 6) & HF- (n = 7) rats 6 weeks post-MI. (B) Western blot (left panel) & quantification (right panel) of O-GlcNAcylated proteins levels measured in proteins extracted from LVs of isolated perfused sham- & HF-rat hearts treated or not with 100 μM thiamet G for 2 h (n = 7 in each group). (C) Western blots (upper panel) & quantification (lower panel) of total desmin levels in the same samples. (D) Phosphorylation profiles of desmin were analyzed in the same samples by Phos-tag™ gel. Graphs show mean ± SEM values expressed in arbitrary units (A.U.). The positions of molecular weight are indicated as kilodalton (kDa) on the left. *P < 0.05; ** < 0.01. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/30344511), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu, Rt, Po, Bv, Dr, Fi, Ha, Pm, V-Vi, Xp
• Applications: WB, ChIP, DB, ELISA, Flow, ICC/IF, IHC, IP, ChIP
• Clone: RL2
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

O-GlcNAc Antibody (RL2) - BSA Free Summary

• Immunogen: Pore complex-lamina fraction purified from rat liver nuclear envelopes.
• Localization: Cytoplasmic and Nuclear
• Specificity: Detects nuclear pore complex (NPC), cytoplasmic and intranuclear O-linked glycoproteins from human, mouse, and rat tissues.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 1:10-1:500
  • Chromatin Immunoprecipitation 1:10 - 1:500. Use reported in scientific literature (PMID 20404350)
  • Dot Blot 1:800
  • ELISA 1:100 - 1:2000. Use reported in scientific literature (PMID 12029848)
  • Flow Cytometry 1:10 - 1:1000
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry 1:10 - 1:500
  • Immunohistochemistry-Paraffin 1:200
  • Immunoprecipitation 1:10 - 1:500
  • Western Blot 1:1000
• Control:

  O-GlcNAc Transferase p110 subunit Overexpression Lysate

Reactivity Notes

Porcine reactivity reported in scientific literature (PMID: 26004176). Xenopus reactivity reported in scientific literature (PMID: 17329255). Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information.

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Protein A purified

Alternate Names for O-GlcNAc Antibody (RL2) - BSA Free

• GlcNAc
• O-linked N-acetylglucosamine

Background

Diffusion of metabolites and small non-nuclear molecules as well as active, mediated import of protein and export of protein and RNA through the nuclear envelope occurs through nuclear pore complexes or NPC's. NPC's contain up to 100 different polypeptides which have a combined mass of about 125 megadaltons. The channel available for passive transport through the NPC is about 9-10 nm in diameter while carrier mediated changes in the NPC result in a ~25 nm channel used for larger, actively transported molecules. Of the 100 polypeptides, at least 8 of these are O-linked N-acetylglycosamine-modified in mammalian cells. All of the mammalian O-linked glycoproteins contain multiple copies of phenylalanine, glycine dipeptide repeats dispersed throughout part of their sequence. Studies indicate that the NPC O-linked glycoproteins have a direct role in nuclear protein import.

Limitations

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.

Publications for O-GlcNAc Antibody (NB300-524)(35)

Publications using NB300-524

• Pelgrom LR, Sergushichev AA, Quik M Protein O-GlcNAcylation and low glycolysis underpin Th2 polarization by dendritic cells Book 2022-01-01 (WB, Mouse)
• Czajewski I, Swain B, Xu J et Al. Rescuable sleep and synaptogenesis phenotypes in a Drosophila model of O-GlcNAc transferase intellectual disability Elife 2024-11-13 [PMID: 39535175]
• Florence Authier, Nina Ondruskova, Andrew T Ferenbach, Alison McNeilly, Daan M F van Aalten Neurodevelopmental defects in a mouse model of O-GlcNAc transferase intellectual disability. Disease models & mechanisms 2024-04-03 [PMID: 38566589]
• Omelková M, Fenger CD, Murray M et al. An O-GlcNAc transferase pathogenic variant linked to intellectual disability affects pluripotent stem cell self-renewal Disease models & mechanisms 2023-06-01 [PMID: 37334838] (WB, Mouse)
• Feng Z, Wang T, Sun Y et al. Sulforaphane suppresses paraquat-induced oxidative damage in bovine�in vitro-matured oocytes through Nrf2 transduction pathway Ecotoxicology and environmental safety 2023-04-01 [PMID: 36907095] (Western Blot, Bovine)
• Kadosaka T, Watanabe M, Natsui H et al. Empagliflozin attenuates arrhythmogenesis in diabetic cardiomyopathy by normalizing intracellular Ca2+ handling in ventricular cardiomyocytes American journal of physiology. Heart and circulatory physiology 2023-03-01 [PMID: 36607794]
• Kumar A, Yarosz EL, Andren A et al. NKT cells adopt a glutamine-addicted phenotype to regulate their homeostasis and function Cell reports 2022-10-25 [PMID: 36288696]
• Zhu WZ, Palazzo T, Zhou M et al. First comprehensive identification of cardiac proteins with putative increased O-GlcNAc levels during pressure overload hypertrophy PloS one 2022-10-26 [PMID: 36288343] (IP)
• Authier F, Attianese B, Bartual S et al. Intellectual disability and neurogenesis defects associated with increased turnover of an O-GlcNAcase variant medRxiv 2023-11-24 (WB)
• Jo R, Shibata H, Kurihara I et al. Mechanisms of mineralocorticoid receptor-associated hypertension in diabetes mellitus: the role of O-GlcNAc modification Hypertension research : official journal of the Japanese Society of Hypertension 2022-10-14 [PMID: 36229526]
• Murray M, Davidson L, Ferenbach A et al. Neuroectoderm phenotypes in a human stem cell model of O-GlcNAc transferase intellectual disability bioRxiv 2023-09-21 (WB, Human)
  
  Details:
  1:1000 dilution
• Authier F, Ondruskova N, Ferenbach A et al. Neurodevelopmental defects in a mouse model of O-GlcNAc transferase intellectual disability bioRxiv 2023-08-24 (WB, Mouse)
• Ise H, Araki Y, Song I, Akatsuka G N-acetylglucosamine-bearing polymers mimicking O-GlcNAc-modified proteins elicit anti-fibrotic activities in myofibroblasts and activated stellate cells Glycobiology 2022-10-03 [PMID: 36190502]
• Shi Q, Shen Q, Liu Y et Al. Increased glucose metabolism in TAMs fuels O-GlcNAcylation of lysosomal Cathepsin B to promote cancer metastasis and chemoresistance Cancer Cell 2022-10-13 [PMID: 36084651]
• Wang TF, Feng ZQ, Sun YW et al. Disruption of O-GlcNAcylation Homeostasis Induced Ovarian Granulosa Cell Injury in Bovine International journal of molecular sciences 2022-07-15 [PMID: 35887161]
• Guilloteau E, COLL P, LU Z Et al. In Utero Exposure to Simulated Complex Urban Air Pollution Disturbs Intestinal Suckling-to-Weaning Transition in Mice Part Fibre Toxicol 2022-06-15 [PMID: 35706036]
• Raab S, Very N, DuchEne B et al. Evaluation of the expression of fatty acid synthase and O GlcNAc transferase in patients with liver cancer by exploration of transcriptome databases and experimental approaches Oncology Letters 2022-02-03 [PMID: 35242233] (IF/IHC, Human)
• Akihito Ishikita, Shouji Matsushima, Soichiro Ikeda, Kosuke Okabe, Ryohei Nishimura, Tomonori Tadokoro, Nobuyuki Enzan, Taishi Yamamoto, Masashi Sada, Yoshitomo Tsutsui, Ryo Miyake, Masataka Ikeda, Tomomi Ide, Shintaro Kinugawa, Hiroyuki Tsutsui GFAT2 mediates cardiac hypertrophy through HBP-O-GlcNAcylation-Akt pathway iScience 2021-12-17 [PMID: 34934932]
• Ogawa Y, Imamoto N Methods to separate nuclear soluble fractions reflecting localizations in living cells iScience 2021-11-01 [PMID: 34934922] (ICC/IF, Human)
• Song T, Zou Q, Yan Y et al. DOT1L O-GlcNAcylation promotes its protein stability and MLL-fusion leukemia cell proliferation Cell reports 2021-09-21 [PMID: 34551297] (WB)
• Muha V, Authier F, Szoke-Kovacs Z et al. Loss of O-GlcNAcase catalytic activity leads to defects in mouse embryogenesis The Journal of biological chemistry 2021-02-18 [PMID: 33610549] (WB, Mouse)
• Murakami K, Kurotaki D, Kawase W, et al. OGT Regulates Hematopoietic Stem Cell Maintenance via PINK1-Dependent Mitophagy Cell reports 2021-01-05 [PMID: 33406421] (FLOW)
• Martinez-Fabregas J, Wang L, Pohler E, Cozzani A CDK8 fine-tunes IL-6 transcriptional activities by limiting STAT3 resident time at the gene loci Cell Rep 2020-12-28 [PMID: 33357429]
• Decourcelle A, Very N, Djouina M et al. O-GlcNAcylation Links Nutrition to the Epigenetic Downregulation of UNC5A during Colon Carcinogenesis Cancers (Basel) 2020-10-28 [PMID: 33126652] (IHC-P, Mouse)
• Nomura A, Yokoe S, Tomoda K et al. Fluctuation in O-GlcNAcylation inactivates STIM1 to reduce store-operated calcium ion entry via downregulation of Ser621 phosphorylation J Biol Chem 2020-10-06 [PMID: 33023909] (WB, Human)
• Mercier T, Bouvet M, Dubois-Deruy E et al. Interplay Between Phosphorylation and O-GlcNAcylation of Sarcomeric Proteins in Ischemic Heart Failure. Front Endocrinol (Lausanne). 2018-10-05 [PMID: 30344511] (WB, Rat)
• Taub DG, Awal MR, Gabel CV., et Al. O-GlcNAc Signaling Orchestrates the Regenerative Response to Neuronal Injury in Caenorhabditis elegans Cell Rep 2018-08-23 [PMID: 30134155]
• Taparra K, Wang H, Malek R et al. O-GlcNAcylation is required for mutant KRAS-induced lung tumorigenesis J. Clin. Invest. 2018-08-21 [PMID: 30130254] (ICC/IF, Mouse)
• Hugo SE, Schlegel A., , et Al. A Genetic Model to Study Increased Hexosamine Biosynthetic Flux Endocrinology 2017-06-06 [PMID: 28582574]
• Swamy M, Pathak S, Grzes K et al. Glucose and glutamine fuel protein O-GlcNAcylation to control T cell self-renewal and malignancy. Nat Immunol. 2016-06-17 [PMID: 27111141] (FLOW, Mouse)
  
  Details:
  This publication used the AF647 conjugated form of this antibody (NB300-524AF647)
• Stateva SR, Villalobo A. O-GlcNAcylation of the human epidermal growth factor receptor Org. Biomol. Chem. 2015-06-25 [PMID: 26108188] (WB, Human)
• Shibutani M, Mori T, Miyano T, Miyake M. Removal of O-GlcNAcylation is important for pig preimplantation development J. Reprod. Dev. 2015-05-22 [PMID: 26004176] (ICC/IF, Porcine)
• Wagner B Dias, Win D Cheung, Gerald W Hart. O-GlcNAcylation of kinases, . Biochemical and Biophysical Research Communications, 10.1016/j.bbrc.2012.04.124. 2012-01-01 [PMID: 22564745] (WB, Human)
• Czajewski I, McDowall L, Ferenbach A Et al Rescuable sleep and synaptogenesis phenotypes in a Drosophila model of O-GlcNAc transferase intellectual disability bioRxiv 2023-06-30 (WB)
• Itkonen HM, Minner S, Guldvik IJ et al. O-GlcNAc transferase integrates metabolic pathways to regulate the stability of c-MYC in human prostate cancer. Cancer Res 2013-05-29 [PMID: 23720054] (IHC-P, Human)

Reviews for O-GlcNAc Antibody (NB300-524) (2)

Average Rating: 4

(Based on 2 reviews)

We have 2 reviews tested in 1 species: Mouse.

reviewed by: Maria Pravata WB Mouse 04/17/2020

Summary

• Application: Western Blot
• Sample Tested: Mouse Embryonic Stem Cells
• Species: Mouse
• Lot: NBP2-22218

reviewed by: Veronica Pravata WB Mouse 01/16/2020

Summary

• Application: Western Blot
• Sample Tested: Mouse Embryonic Stem Cells
• Species: Mouse
• Lot: A-3

Comments

• Comments: Nitrocellulose membrane, Blocked in 5% BSA in PBS-T, 1:1000 ab overnight at 4 degrees.

FAQs for O-GlcNAc Antibody (NB300-524). (Showing 1 - 1 of 1 FAQs).

1. Is it possible to get your mAb without BSA or sodium azide? I am interested in NB300-614 and NB300-524 .
   • These antibodies are supplied in Sodium Azide, but if you are interested, we do provide kits to clean up the antibodies. Here is the link to our AbSelect Antibody Purification Kits .