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Western blot
ELISAAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
- Immunohistochemistry [1]
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- Product number
- MA5-33118 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- BAG3 Recombinant Rabbit Monoclonal Antibody (3D8)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 3D8
- Vial size
- 100 µL
- Concentration
- 0.25 mg/mL
- Storage
- -20°C or -80°C if preferred
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-BAG3 Recombinant Rabbit Monoclonal Antibody (Product # MA5-33118) and a 74 kDa band corresponding to BAG family molecular chaperone regulator 3 was observed across the first three cell lines. Whole cell extracts (30 µg lysate) of HT-1080 (Lane 1), U-87 MG (Lane 2), DU 145 (Lane 3), SW480 (Lane 4), HEL 92.1.7 (Lane 5), K-562 (Lane 6), Jurkat (Lane 7) 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 (1:3000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:10000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of BAG3 using a BAG3 Monoclonal antibody (Product # MA5-33118) at a concentration of 0.73 µg/mL. Positive WB detected in: Hela whole cell lysate, 293 whole cell lysate, A549 whole cell lysate, HepG2 whole cell lysate, U87 whole cell lysate. A secondary Goat polyclonal antibody to rabbit IgG was applied at a 1:50,000 dilution. Observed band size: 80 kDa.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of BAG3 in paraffin embedded human breast cancer using a BAG3 monoclonal antibody (Product # MA5-33118) at a dilution of 1:73.75. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.
