This gene encodes the alpha subunit of glucosidase II and a member of the glycosyl hydrolase 31 family of proteins. The heterodimeric enzyme glucosidase II plays a role in protein folding and quality control by cleaving glucose residues from immature glycoproteins in the endoplasmic reticulum. Expression of the encoded protein is elevated in lung tumor tissue and in response to UV irradiation. Mutations in this gene cause autosomal-dominant polycystic kidney and liver disease. atalytic subunit of glucosidase II that cleaves sequentially the 2 innermost alpha-1,3-linked glucose residues from the Glc(2)Man(9)GlcNAc(2) oligosaccharide precursor of immature glycoproteins. Required for PKD1/Polycystin-1 and PKD2/Polycystin-2 maturation and localization to the cell surface and cilia.
Background References
1. Pelletier M.F., Marcil A., Sevigny G., Jakob C.A., Tessier D.C., Chevet E., Menard R., Bergeron J.J.M., Thomas D.Y. The heterodimeric structure of glucosidase II is required for its activity, solubility, and localization in vivo. Glycobiology 10:815-827 (2000)
2. Porath B., Gainullin V.G., Cornec-Le Gall E., Dillinger E.K., Heyer C.M., Hopp K., Edwards M.E., Madsen C.D., et al. Mutations in GANAB, encoding the glucosidase IIalpha subunit, cause autosomal-dominant polycystic kidney and liver disease. Am. J. Hum. Genet. 98:1193-1207 (2016)
Western blot analysis of GANAB on different lysates with Rabbit anti-GANAB antibody (HA721695) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.
Lane 1: HeLa cell lysate Lane 2: HepG2 cell lysate Lane 3: HEK-293 cell lysate Lane 4: Raji cell lysate Lane 5: NIH/3T3 cell lysate Lane 6: C2C12 cell lysate Lane 7: PC-12 cell lysate
Lysates/proteins at 15 µg/Lane.
Predicted band size: 107 kDa Observed band size: 107 kDa
Exposure time: 1 minute 21 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721695) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunofluorescence analysis of paraffin-embedded human thyroid tissue labeling GANAB with Rabbit anti-GANAB antibody (HA721695) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA721695, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
Western blot analysis of GANAB on different lysates with Rabbit anti-GANAB antibody (HA721695) at 1/1,000 dilution.
Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: HepG2 cell lysate (20 µg/Lane) Lane 3: LoVo cell lysate (20 µg/Lane) Lane 4: HEK-293 cell lysate (20 µg/Lane) Lane 5: U-2 OS cell lysate (20 µg/Lane) Lane 6: K-562 cell lysate (20 µg/Lane) Lane 7: Jurkat cell lysate (20 µg/Lane) Lane 8: HL-60 cell lysate (20 µg/Lane) Lane 9: Raji cell lysate (20 µg/Lane) Lane 10: NIH/3T3 cell lysate (20 µg/Lane) Lane 11: C2C12 cell lysate (20 µg/Lane) Lane 12: RAW264.7 cell lysate (20 µg/Lane) Lane 13: Mouse spleen tissue lysate (40 µg/Lane)
Predicted band size: 107 kDa Observed band size: 107 kDa
Exposure time: 1 minute 2 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721695) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunofluorescence analysis of paraffin-embedded mouse epididymis tissue labeling GANAB with Rabbit anti-GANAB antibody (HA721695) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA721695, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
Western blot analysis of GANAB on different lysates with Rabbit anti-GANAB antibody (HA721695) at 1/1,000 dilution.
Lane 1: PC-12 cell lysate (20 µg/Lane) Lane 2: Mouse placenta tissue lysate (40 µg/Lane) Lane 3: Rat brain tissue lysate (40 µg/Lane) Lane 4: Human liver tissue lysate (40 µg/Lane)
Predicted band size: 107 kDa Observed band size: 107 kDa
Exposure time: 3 minutes 10 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721695) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Knockdown (KD)
Western blot analysis of GANAB on different lysates with Rabbit anti-GANAB antibody (HA721695) at 1/5,000 dilution.
Lane 1: HeLa-si NT cell lysate Lane 2: HeLa-si GANAB cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 107 kDa Observed band size: 107 kDa
Exposure time: 3 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721695) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-GANAB antibody (HA721695) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721695) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-GANAB antibody (HA721695) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721695) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human thyroid tissue with Rabbit anti-GANAB antibody (HA721695) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721695) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded mouse epididymis tissue with Rabbit anti-GANAB antibody (HA721695) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721695) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
GANAB was immunoprecipitated in 0.2mg HeLa cell lysate with HA721695 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA721695 at 1/1,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input) Lane 2: Rabbit IgG instead of HA721695 in HeLa cell lysate Lane 3: HA721695 IP in HeLa cell lysate