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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [2]
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- Product number
- 14-9933-80 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Caspase 7 Monoclonal Antibody (20F3), eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The monoclonal antibody 20F3 recognizes human caspase-7, also known as mch3 and ICE-LAP3. It is a member of the cysteine-requiring aspartate protease family, which plays critical roles in cell suicide known as apoptosis. Along with caspases 3 and 7, this member is categorized as an effector or executioner caspase. It must first be activated via cleavage by capase 3, 6, or 8 or by granzyme B. As an effector molecule, caspase-7 is responsible for the demolition phase of apoptosis in that it cleaves PARP and other molecules also cleaved by caspase-3. This monoclonal antibody can be used to identify both the full-length (p35) and active form (p20) of caspase-7 under reducing conditions by SDS-PAGE. Applications Reported: This 20F3 antibody has been reported for use in immunoblotting (WB). Applications Tested: This 20F3 antibody has been tested by immunoblot analysis of reduced Jurkat cell lysates. This can be used at 1-5 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. The presence of a nonspecific band at approximately 58 kDa may be present in some lysates. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human
- Host
- Rat
- Isotype
- IgG
- Antibody clone number
- 20F3
- Vial size
- 25 µg
- Concentration
- 0.5 mg/mL
- Storage
- 4° C
Submitted references Engineered hybrid dimers: tracking the activation pathway of caspase-7.
Denault JB, Békés M, Scott FL, Sexton KM, Bogyo M, Salvesen GS
Molecular cell 2006 Aug;23(4):523-33
Molecular cell 2006 Aug;23(4):523-33
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Jurkat cell lysates reduced with DTT were resolved by SDS-PAGE then immunoblotted with 2 µg/mL of Anti-Human Caspase 7 Purified. Bands were visualized using Anti-Rat IgG HRP.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Caspase 7 Monoclonal Antibody (20F3), eBioscience™ (Product # 14-9933-80) and a 35 kDa band corresponding to Caspase-7 was observed in cell line tested and decreased upon Etoposide treatment due to cleavage of the full length Caspase-7. Whole cell extracts (30 µg lysate) of Jurkat (Lane 1) and Jurkat treated with Etoposide (25 µM for 16hr) (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 (0.5 µg/mL) and detected by chemiluminescence with F(ab2-Rabbit anti-Rat IgG (H+L Secondary Antibody, HRP (Product # PA1-29927,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
