SMAC/Diablo Antibody (78-1-118) - BSA Free
Novus Biologicals | Catalog # NB500-213
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Key Product Details
Species Reactivity
Validated:
Human, Mouse, Rat
Cited:
Mouse, Rat
Applications
Validated:
Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation
Cited:
Western Blot, Immunocytochemistry/ Immunofluorescence
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG1 Clone # 78-1-118
Format
BSA Free
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Product Specifications
Immunogen
Recombinant mature human Smac (lacks amino acids 1-55 present in Smac precursor). [UniProt# Q9NR28]
Localization
Mitochondrial but released into the cytosol when cells undergo apoptosis.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG1
Theoretical MW
25 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Scientific Data Images for SMAC/Diablo Antibody (78-1-118) - BSA Free
Western Blot: SMAC/Diablo Antibody (78-1-118) [NB500-213]
Western Blot: Smac Antibody (78-1-118) [NB500-213] - HeLa whole cell extract lysates.Applications for SMAC/Diablo Antibody (78-1-118) - BSA Free
Application
Recommended Usage
Immunoprecipitation
1:10 - 1:500
Western Blot
1:500
Application Notes
This Smac (78-1-118) antibody is useful for Immunoprecipitation and Western blot, where a band is seen at ~25 kDa. For better results, IP prior to Western blot is recommended. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Reviewed Applications
Read 4 reviews rated 4.8 using NB500-213 in the following applications:
Formulation, Preparation, and Storage
Purification
Unpurified
Formulation
Ascites
Format
BSA Free
Preservative
0.1% Sodium Azide
Concentration
This product is unpurified. The exact concentration of antibody is not quantifiable.
Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
Background: SMAC/Diablo
Long Name
Second Mitochondria-derived Activator of Caspase
Alternate Names
Diablo
Gene Symbol
DIABLO
UniProt
Additional SMAC/Diablo Products
Product Documents for SMAC/Diablo Antibody (78-1-118) - BSA Free
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Product Specific Notices for SMAC/Diablo Antibody (78-1-118) - BSA Free
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
Related Research Areas
Citations for SMAC/Diablo Antibody (78-1-118) - BSA Free
Customer Reviews for SMAC/Diablo Antibody (78-1-118) - BSA Free (4)
4.8 out of 5
4 Customer Ratings
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Application: Western BlotSample Tested: H9c2 whole cell lysateSpecies: RatVerified Customer | Posted 02/07/2022SMAC
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Application: Western BlotSample Tested: AC!6 and AC16Species: HumanVerified Customer | Posted 04/20/2021SMAC
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Application: Western BlotSample Tested: AC16Species: MouseVerified Customer | Posted 05/15/2019SMAC/DIABLO
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Application: Western BlotSample Tested: AC16Species: HumanVerified Customer | Posted 05/06/2019DIABLO
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Protocols
View specific protocols for SMAC/Diablo Antibody (78-1-118) - BSA Free (NB500-213):
SMAC/Diablo Antibody (78-1-118):
Procedure Guide for NB 500-213 Monoclonal anti-Survivin (clone 58.22)
Immunoprecipitation Procedure
1) Lyse cells plated in a 60mm dish:
a) 300 ul CHAPS buffer [50mM Tris-HCl, pH 7.5/50mM NaCl/1mM EDTA/1% NP-40/0.1% CHAPS/1mM NaVO4/1mM PMSF]
b) rock for 20 minutes at 4C
2) Harvest lysate and spin down the insoluble material at 14K rpm.
3) Collect soluble fraction.
4) Pre-clear lysate with 40 ul of 50:50 slurry of Protein A beads, rocking for 1 hour at 4C.
5) Spin down beads at 2K rpm, at 4C.
6) Collect pre-cleared lysate.
7) Incubate lysate with 5-7ug of anti-Survivin (NB 500-213) overnight, rocking at 4C.
8) Add 50 ul of Protein A 50:50 slurry for 2 hours, rocking at 4C.
9) Wash beads with 200 ul of CHAPS buffer, three times.
10) Denature immune complex by adding 2x Sample Buffer, containing 2-ME.
11) Boil for 10 minutes and load onto an SDS-gel.
Procedure Guide for NB 500-213 Monoclonal anti-Survivin (clone 58.22)
Immunoprecipitation Procedure
1) Lyse cells plated in a 60mm dish:
a) 300 ul CHAPS buffer [50mM Tris-HCl, pH 7.5/50mM NaCl/1mM EDTA/1% NP-40/0.1% CHAPS/1mM NaVO4/1mM PMSF]
b) rock for 20 minutes at 4C
2) Harvest lysate and spin down the insoluble material at 14K rpm.
3) Collect soluble fraction.
4) Pre-clear lysate with 40 ul of 50:50 slurry of Protein A beads, rocking for 1 hour at 4C.
5) Spin down beads at 2K rpm, at 4C.
6) Collect pre-cleared lysate.
7) Incubate lysate with 5-7ug of anti-Survivin (NB 500-213) overnight, rocking at 4C.
8) Add 50 ul of Protein A 50:50 slurry for 2 hours, rocking at 4C.
9) Wash beads with 200 ul of CHAPS buffer, three times.
10) Denature immune complex by adding 2x Sample Buffer, containing 2-ME.
11) Boil for 10 minutes and load onto an SDS-gel.
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunoprecipitation Protocol
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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