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Product Info Summary
| SKU: | A00080-5 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Caspase-9/CASP9 Antibody Picoband®
SKU/Catalog Number
A00080-5
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Caspase-9/CASP9 Antibody Picoband® catalog # A00080-5. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-Caspase-9/CASP9 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00080-5)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human Caspase-9/CASP9 recombinant protein (Position: E3-K410).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
A00080-5 is reactive to CASP9 in Human
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
46 kDa
Calculated molecular weight
15816 MW
Background of Caspase-9
CASP9 is also known as MCH6 or APAF3. This gene encodes a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. This protein can undergo autoproteolytic processing and activation by the apoptosome, a protein complex of cytochrome c and the apoptotic peptidase activating factor 1; this step is thought to be one of the earliest in the caspase activation cascade. This protein is thought to play a central role in apoptosis and to be a tumor suppressor. Alternative splicing results in multiple transcript variants.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A00080-5 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human Hela whole cell, human A549 whole cell
IHC: human lung cancer tissue
ICC/IF: HELA cell
FCM: U937 cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of Caspase-9/CASP9 using anti-Caspase-9/CASP9 antibody (A00080-5).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human A549 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Caspase-9/CASP9 antigen affinity purified polyclonal antibody (Catalog # A00080-5) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Caspase-9/CASP9 at approximately 46KD. The expected band size for Caspase-9/CASP9 is at 46KD.
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Figure 2. IHC analysis of Caspase-9/CASP9 using anti-Caspase-9/CASP9 antibody (A00080-5).
Caspase-9/CASP9 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-Caspase-9/CASP9 Antibody (A00080-5) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
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Figure 3. IF analysis of Caspase-9/CASP9 using anti-Caspase-9/CASP9 antibody (A00080-5).
Caspase-9/CASP9 was detected in immunocytochemical section of HELA cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-Caspase-9/CASP9 Antibody (A00080-5) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Figure 4. Flow Cytometry analysis of U937 cells using anti-Caspase-9/CASP9 antibody (A00080-5).
Overlay histogram showing U937 cells stained with A00080-5 (Blue line).To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Caspase-9/CASP9 Antibody (A00080-5, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For CASP9 (Source: Uniprot.org, NCBI)
Gene Name
CASP9
Full Name
Caspase-9
Weight
15816 MW
Superfamily
peptidase C14A family
Alternative Names
Interleukin-13;IL-13;IL13;NC30; CASP9 APAF-3, APAF3, ICE-LAP6, MCH6, PPP1R56 caspase 9 caspase-9|ICE-like apoptotic protease 6|apoptotic protease MCH-6|apoptotic protease activating factor 3|caspase 9, apoptosis-related cysteine peptidase|protein phosphatase 1, regulatory subunit 56
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on CASP9, check out the CASP9 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for CASP9: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-Caspase-9/CASP9 Antibody Picoband® (A00080-5)
Hello CJ!
A00080-5 has been cited in 1 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Li QY,Hou CZ,Yang LP,Chu XL,Wang Y,Zhang P,Zhao Y. Study on the Mechanism of Ginseng in the Treatment of Lung Adenocarcinoma Based on Network Pharmacology. Evid Based Complement Alternat Med.2020 Jul 31;2020:2658795. doi:10.1155/2020/2658795.PMID:32802118
Species: Human
A00080-5 usage in article: APP:ICC, SAMPLE:A549 CELL, DILUTION:NA
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