Explore
Validate
Learn
Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
- Other assay [1]
Submit
Validation data
Reference
Comment
Report error
- Product number
- PA5-27940 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- GNAI3 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, H1299, HeLaS3, HepG2, Molt-4, Raji, mouse brain.
- Concentration
- 0.94 mg/mL
Submitted references Downregulation of GNAI3 Promotes the Pathogenesis of Methionine/Choline-Deficient Diet-Induced Nonalcoholic Fatty Liver Disease.
Zhu H, Ge K, Lu J, Jia C
Gut and liver 2020 Jul 15;14(4):492-499
Gut and liver 2020 Jul 15;14(4):492-499
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using GNAI3 Polyclonal Antibody (Product # PA5-27940). Sample (30 µg whole cell lysate). A: A431. B: MOLT4. C: Raji. 7.5% SDS PAGE. GNAI3 Polyclonal Antibody (Product # PA5-27940) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using GNAI3 Polyclonal Antibody (Product # PA5-27940). Sample (50 µg of whole cell lysate). Lane A: Mouse brain. 10% SDS PAGE. GNAI3 Polyclonal Antibody (Product # PA5-27940) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of GNAI3 was achieved by transfecting A-431 cells with GNAI3 specific siRNAs (Silencer® select Product # s5878). Western blot analysis (Fig. a) was performed whole cell extracts from the GNAI3 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with GNAI3 Polyclonal Antibody (Product # PA5-27940, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25 µg/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 GNAI3.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-GNAI3 Rabbit Polyclonal Antibody (Product # PA5-27940) and a 40kDa band corresponding to GNAI3 was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of A-431 (Lane 1), SH-SY5Y (Lane 2), U-87 MG (Lane 3), U-2 OS (Lane 4), MOLT-4 (Lane 5), Mouse Brain (Lane 6) and Mouse Lung (Lane 7) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of GNAI3 was performed using 70% confluent log phase A-431 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 GNAI3 Polyclonal Antibody (Product # PA5-27940) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear and 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)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded SAS xenograft, using G protein alpha Inhibitor 3 (Product # PA5-27940) antibody at 1:500 dilution. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Fig. 3 Construction of GNAI3 KD HepG2 cells and the effect of GNAI3 KD on cellular lipid accumulation. (A) Quantitative reverse transcription polymerase chain reaction analysis of GNAI3 mRNA expression in HepG2 cells transfected with lentiviral vectors carrying GNAI3 shRNA. (B) Western blotting analysis of GNAI3 protein expression levels in GNAI3 KD and control HepG2 cells. (C) Oil Red O staining of control, GNAI3 KD and rescued HepG2 cells after free fatty acid treatment. Positive staining for Oil Red O indicates intracellular lipid accumulation. Quantification of Oil Red O staining is shown in the right panel. The bar graphs show the mean+-standard error of the mean. p
