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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
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Validation data
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- Product number
- 710856 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Golgi Protein 58K Recombinant Polyclonal Antibody (17 HCLC)
- Antibody type
- Polyclonal
- Antigen
- Other
- Description
- This antibody is predicted to react with Monkey, Pig and Bovine
- Antibody clone number
- 17 HCLC
- Concentration
- 0.5 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Golgi Protein 58k Recombinant Polyclonal Antibody (Product # 710856) and a 58kDa band corresponding to Golgi Protein 58k was observed in Rat Liver but not in Rat Brain which is reported to be negative. Tissue lysates (30ug) of Rat Liver (Lane 1) and Rat Brain (Lane 2) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein 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 (0.5ug/ml) 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). An uncharacterized band was also observed at ~80kDa.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence was performed on methanol fixed HepG2 cells for detection of endogenous Golgi protein 58K using Anti-Golgi protein 58K Recombinant Rabbit Polyclonal Antibody (Product # 710856, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of Golgi protein 58K protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) Cytoskeleton was stained with alpha-tubulin Monoclonal Antibody (Product # 32-2500, 1 µg/mL) followed by Goat anti-Mouse IgG Secondary Antibody, Alexa Fluor® 594 conjugate (Product # A-11032, 1:400). Panel d) is a composite image of Panels a, b and c clearly demonstrating staining for Golgi protein 58K in the perinuclear area, indicating localization in the golgi apparatus. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
