O-GlcNAc Antibody (RL2) - Azide and BSA Free

Western Blot: O-GlcNAc Antibody (RL2) - Azide and BSA Free [NBP2-80892] - 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. Image from the standard format of this antibody.

Immunocytochemistry/Immunofluorescence: O-GlcNAc Antibody (RL2) - Azide and BSA Free [NBP2-80892] - 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. Image from the standard format of this antibody.

Immunohistochemistry: O-GlcNAc Antibody (RL2) - Azide and BSA Free [NBP2-80892] - 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. Image from the standard format of

Flow (Intracellular): O-GlcNAc Antibody (RL2) - Azide and BSA Free [NBP2-80892] - 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 directly conjugated to Alexa Fluor 647.

Immunocytochemistry/Immunofluorescence: O-GlcNAc Antibody (RL2) - Azide and BSA Free [NBP2-80892] - 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. Image from the standard format of this antibody.

Flow Cytometry: O-GlcNAc Antibody (RL2) - Azide and BSA Free [NBP2-80892] - 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 directly conjugated to Alexa Fluor 647.

Flow Cytometry: O-GlcNAc Antibody (RL2) - Azide and BSA Free [NBP2-80892] - 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 directly conjugated to phycoerythrin.

Flow Cytometry: O-GlcNAc Antibody (RL2) - Azide and BSA Free [NBP2-80892] - 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 directly conjugated to Alexa Fluor 647.

Flow Cytometry: O-GlcNAc Antibody (RL2) - Azide and BSA Free [NBP2-80892] - 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 directly conjugated to Alexa Fluor 647.

Flow Cytometry: O-GlcNAc Antibody (RL2) - Azide and BSA Free [NBP2-80892] - 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 directly conjugated to Alexa Fluor 647.

Flow Cytometry: O-GlcNAc Antibody (RL2) - Azide and BSA Free [NBP2-80892] - 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 directly conjugated to Alexa Fluor 647.

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

O-GlcNAc Antibody (RL2) - Azide and BSA Free Summary

• Immunogen: Pore complex-lamina fraction purified from rat liver nuclear envelopes.
• Localization: Cytoplasmic and Nuclear
• 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)
  • CyTOF-ready
  • Dot Blot 1:800
  • ELISA 1:100 - 1:2000,. Use reported in scientific literature (PMID 12029848)
  • Flow (Intracellular)
  • 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

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: No Preservative
• Concentration: 1.0 mg/ml
• Purity: Protein A purified

Alternate Names for O-GlcNAc Antibody (RL2) - Azide and 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.