MA1-038

antibody from Invitrogen Antibodies

Targeting: OGT FLJ23071, HRNT1, MGC22921, O-GLCNAC, OGT1

Western blot (Data presented on provider website)

Immunoprecipitation (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-038

Provider

Invitrogen Antibodies

Product name

Anti-O-linked N-acetylglucosamine (O-GlcNAc) Monoclonal Antibody (18B10.C7)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

MA1-038 has been successfully used in Western blot and immunoprecipitation applications with human samples. In Western blot, MA1-038 detects proteins with O-GlcNAc post-translational modification, and therefore will detect multiple specific bands.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

18B10.C7

Vial size

100 µg

Concentration

1 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

O-GlcNAcylation of G6PD promotes the pentose phosphate pathway and tumor growth.

Rao X, Duan X, Mao W, Li X, Li Z, Li Q, Zheng Z, Xu H, Chen M, Wang PG, Wang Y, Shen B, Yi W
Nature communications 2015 Sep 24;6:8468

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The intracellular fate of zonula occludens 2 is regulated by the phosphorylation of SR repeats and the phosphorylation/O-GlcNAcylation of S257.

Quiros M, Alarcón L, Ponce A, Giannakouros T, González-Mariscal L
Molecular biology of the cell 2013 Aug;24(16):2528-43

Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation of Glycogen Synthase was performed on HepG2 cells, glucose and serum starved for 36 hours. The antigen:antibody complex was formed by binding 500 µg whole cell lysate with 5 µg of mouse monoclonal antibody recognizing O-GlcNAc (Product # MA1-038) overnight on a rocking platform at 4°C. The immune-complex was then captured on 50 µL Protein A/G Plus Agarose (Product # 20424). The beads were washed to remove non-bound material and a low-pH elution buffer was used to dissociate bound antigen from the antibody-bound bead. Samples were then resolved on a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to PVDF membrane and blocked with 5% BSA/TBS-0.1%Tween for at least 1 hour. Membranes were then probed with a mouse monoclonal antibody recognizing Glycogen Synthase (Product # PA5-17459) at a dilution of 1:1000 overnight rotating at 4°C. Membranes were then washed in TBST and probed with Clean-blot IP detection reagent (Product # 21230) at a dilution of 1:2000 for at least one hour. Membranes were washed and chemiluminescent detection was performed using Super Signal West Pico (Product # 34087). The image is a Western blot with antibody Product # PA5-17459 showing enrichment of glycogen synthase by O-GlcNAc precipitation. Lane 1 indicates 25 µg of HepG2 whole cell lysate input; Lane 2 indicates the IP elution.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation of Glycogen Synthase was performed on HepG2 cells, glucose and serum starved for 36 hours. The antigen:antibody complex was formed by binding 500 µg whole cell lysate with 5 µg of mouse monoclonal antibody recognizing O-GlcNAc (Product # MA1-038) overnight on a rocking platform at 4°C. The immune-complex was then captured on 50 µL Protein A/G Plus Agarose (Product # 20424). The beads were washed to remove non-bound material and a low-pH elution buffer was used to dissociate bound antigen from the antibody-bound bead. Samples were then resolved on a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to PVDF membrane and blocked with 5% BSA/TBS-0.1%Tween for at least 1 hour. Membranes were then probed with a mouse monoclonal antibody recognizing Glycogen Synthase (Product # PA5-17459) at a dilution of 1:1000 overnight rotating at 4°C. Membranes were then washed in TBST and probed with Clean-blot IP detection reagent (Product # 21230) at a dilution of 1:2000 for at least one hour. Membranes were washed and chemiluminescent detection was performed using Super Signal West Pico (Product # 34087). The image is a Western blot with antibody Product # PA5-17459 showing enrichment of glycogen synthase by O-GlcNAc precipitation. Lane 1 indicates 25 µg of HepG2 whole cell lysate input; Lane 2 indicates the IP elution.