O-GlcNAc Antibody (RL2) - BSA Free

Novus Biologicals | Catalog # NB300-524

Novus Biologicals
100% Guarantee

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Bovine, Drosophila, Fish, Hamster, Primate, Virus, Xenopus

Cited:

Human, Mouse, Rat, Porcine, Bovine

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Chromatin Immunoprecipitation, Chromatin Immunoprecipitation (ChIP), Dot Blot

Cited:

Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # RL2

Format

BSA Free
View all O-GlcNAc Primary Antibodies »

Product Specifications

Immunogen

Pore complex-lamina fraction purified from rat liver nuclear envelopes.

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.

Localization

Cytoplasmic and Nuclear

Specificity

Detects nuclear pore complex (NPC), cytoplasmic and intranuclear O-linked glycoproteins from human, mouse, and rat tissues.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for O-GlcNAc Antibody (RL2) - BSA Free

Immunohistochemistry-Paraffin: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524]

Immunohistochemistry-Paraffin: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524]

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)BSA Free [NB300-524]

Western Blot: O-GlcNAc Antibody (RL2)BSA Free [NB300-524]

O-GlcNAc-Antibody-RL2-Western-Blot-NB300-524-img0016.jpg
Immunocytochemistry/ Immunofluorescence: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524]

Immunocytochemistry/ Immunofluorescence: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524]

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) - BSA Free [NB300-524]

Flow Cytometry: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524]

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)BSA Free [NB300-524]

Western Blot: O-GlcNAc Antibody (RL2)BSA Free [NB300-524]

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) - BSA Free [NB300-524]

Immunocytochemistry/ Immunofluorescence: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524]

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) - BSA Free [NB300-524]

Flow Cytometry: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524]

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) - BSA Free [NB300-524]

Flow Cytometry: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524]

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) - BSA Free [NB300-524]

Flow Cytometry: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524]

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) - BSA Free [NB300-524]

Flow Cytometry: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524]

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) - BSA Free [NB300-524]

Flow Cytometry: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524]

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) - BSA Free [NB300-524]

Flow Cytometry: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524]

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) - BSA Free [NB300-524]

Flow Cytometry: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524]

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) - BSA Free [NB300-524]

Flow Cytometry: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524]

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).
O-GlcNAc Antibody (RL2) - BSA Free

Western Blot: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524] -

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 (https://pubmed.ncbi.nlm.nih.gov/30344511), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
O-GlcNAc Antibody (RL2) - BSA Free

Western Blot: O-GlcNAc Antibody (RL2) - BSA Free [NB300-524] -

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 (https://pubmed.ncbi.nlm.nih.gov/30344511), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for O-GlcNAc Antibody (RL2) - BSA Free

Application
Recommended Usage

Chromatin Immunoprecipitation

1:10 - 1:500. Use reported in scientific literature (PMID 20404350)

Chromatin Immunoprecipitation (ChIP)

1:10-1:500

Dot Blot

1:800

ELISA

1:100 - 1:2000. Use reported in scientific literature (PMID 12029848)

Flow Cytometry

1:10 - 1:1000

Immunohistochemistry

1:10 - 1:500

Immunohistochemistry-Paraffin

1:200

Immunoprecipitation

1:10 - 1:500

Western Blot

1:1000

Reviewed Applications

Read 2 reviews rated 4 using NB300-524 in the following applications:

Flow Cytometry Panel Builder

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: O-GlcNAc

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.

Alternate Names

GlcNAc, O-linked N-acetylglucosamine

Additional O-GlcNAc Products

Product Documents for O-GlcNAc Antibody (RL2) - BSA Free

Certificate of Analysis

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Product Specific Notices for O-GlcNAc Antibody (RL2) - BSA Free

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.

Citations for O-GlcNAc Antibody (RL2) - BSA Free

Customer Reviews for O-GlcNAc Antibody (RL2) - BSA Free (2)

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Customer Images

Showing  1 - 2 of 2 reviews Showing All
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  • O-GlcNAc Antibody (RL2)
    Name: Maria Pravata
    Application: Western Blot
    Sample Tested: Mouse Embryonic Stem Cells
    Species: Mouse
    Verified Customer | Posted 04/17/2020
    Glut2 expression in two different culture conditions. The nitrocellulose membrane was incubated overnight with Glut2 ab at a concentration of 1:1000 in 5% BSA in PBS-Tween (0.01%)
    O-GlcNAc Antibody (RL2) - BSA Free NB300-524
  • O-GlcNAc Antibody (RL2)
    Name: Veronica Pravata
    Application: Western Blot
    Sample Tested: Mouse Embryonic Stem Cells
    Species: Mouse
    Verified Customer | Posted 01/16/2020
    O-GlcNAc Western blot from 20 ug of mESCs lysate
    Nitrocellulose membrane, Blocked in 5% BSA in PBS-T, 1:1000 ab overnight at 4 degrees.
    O-GlcNAc Antibody (RL2) - BSA Free NB300-524

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Protocols

View specific protocols for O-GlcNAc Antibody (RL2) - BSA Free (NB300-524):


Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.


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FAQs for O-GlcNAc Antibody (RL2) - BSA Free

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  • Q: Is it possible to get your mAb without BSA or sodium azide? I am interested in NB300-614 and NB300-524.

    A:

    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 .

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