- PA5-21431 - Invitrogen Antibodies GANAB antibody

Kuribara T, Imagawa A, Hirano M, Ito Y, Totani K
FEBS letters 2020 Jun;594(11):1759-1769

Lamriben L, Oster ME, Tamura T, Tian W, Yang Z, Clausen H, Hebert DN
The Journal of biological chemistry 2018 Sep 7;293(36):13932-13945

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using GANAB Polyclonal Antibody (Product # PA5-21431). Sample (30 µg whole cell lysate). Lane A: 293T. B: A431. C: H1299. D: HeLa S3. E: HepG2. F: MOLT4. G: Raji. 7.5% SDS PAGE. GANAB Polyclonal Antibody (Product # PA5-21431) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using GANAB Polyclonal Antibody (Product # PA5-21431). Sample (50 µg of whole cell lysate). Lane A: Mouse brain. 7.5% SDS PAGE. GANAB Polyclonal Antibody (Product # PA5-21431) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of GANAB was achieved by transfecting HeLa with GANAB specific siRNAs (Silencer® select Product # s23248; s23247). Western blot analysis (Fig. a) was performed using whole cell extracts from the GANAB knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with GANAB Polyclonal Antibody (Product # PA5-21431, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to GANAB.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), A-431 (Lane 2), Hep G2 (Lane 3), MOLT-4 (Lane 4), Raji (Lane 5), NIH/3T3 (Lane 6) and tissue extract of Mouse Liver (Lane 7). The blot was probed with Anti-GANAB Polyclonal Antibody (Product # PA5-21431, 1:3000 dilution) and detected by chemiluminescence using Goat anti Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 109 kDa band corresponding to GANAB was observed across all cell lines and tissue extracts tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of alpha Glucosidase II in methanol-fixed HeLa cells using an Alpha Glucosidase II polyclonal antibody (Product # PA5-21431) (Green) at a 1:500 dilution. Alpha-tubulin filaments were labeled with Product # PA5-29281 (Red) at a 1:2000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Confocal immunofluorescence analysis (Olympus FV10i) of methanol-fixed HeLa, using alpha Glucosidase II antibody (Product # PA5-21431) (Green) at 1:500 dilution. Alpha-tubulin filaments were labeled with (Product # MA1-25054) (Red) at 1:2,000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of GANAB was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with GANAB Rabbit Polyclonal Antibody (Product # PA5-21431) at 5 microgram/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing Cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry (Paraffin) analysis of GANAB was performed in paraffin-embedded human breast carcinoma tissue using GANAB Polyclonal Antibody (Product # PA5-21431) at a dilution of 1:500.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of paraffin-embedded SW480 xenograft , using alpha Glucosidase II (Product # PA5-21431) antibody at 1:500 dilution. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 2 Fig. Relative amounts of glucosidase II and uridine diphosphate-glucose: glycoprotein glucosyltransferase 1 mRNA and protein in obesity model rats. Relative amounts of mRNA of Ganab (glucosidase II alpha-subunit) (A), Prkcsh (glucosidase II beta-subunit) (B), and Uggt1 (C) in Zucker normal (ZN) and Zucker fatty (ZF) rats are represented in the bar graphs. beta2 Microglobulin was used as normalization control. Relative protein quantities of glucosidase II alpha-subunit (GIIalpha) (D), glucosidase II beta-subunit (GIIbeta) (E), and uridine diphosphate-glucose: glycoprotein glucosyltransferase 1 (UGGT1) (F) in ZN and ZF rats are represented in the bar graphs. Typical western blot results are shown below the bar graphs. beta-Actin was used as loading control for normalization of all examined protein quantities. The homogenates were applied at 10 µg protein/lane. Data are expressed as mean +- standard deviation ( n = 3). Statistical significance is indicated by *, **, and *** referring to P < 0.05, 0.01, and 0.001, respectively

PA5-21431