Anti-Caspase-3(P17)/CASP3 Antibody

Product Name

Anti-Caspase-3(P17)/CASP3 Antibody

View all Caspase-3 Antibodies

SKU/Catalog Number

PA1961-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Caspase-3 (P17)/CASP3 Antibody catalog # PA1961-1. Tested in IHC, WB applications. This antibody reacts with Mouse, Rat.

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-3(P17)/CASP3 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1961-1)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence at the N-terminus of mouse Caspase-3(P17), different from the related rat sequence by one amino acid.

*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

PA1961-1 is reactive to CASP3 in Mouse, Rat

Applications

PA1961-1 is guaranteed for IHC, WB Boster Guarantee

Observed Molecular Weight

32 kDa, 19 kDa

Calculated molecular weight

31.608kDa

Background of Caspase-3

CASP3 (Caspase3 Apoptosis-Related Cysteine Protease), also known as YAMA, CPP32 or APOPAIN, is a caspase protein that interacts with caspase 8 and caspase 9. It is encoded by the CASP3 gene. The CASP3 protein is a member of the cysteine-aspartic acid protease (caspase) family. Tiso et al. (1996) used radiation hybrid mapping to localize the CPP32 gene to human chromosome 4q33-q35.1. Fernandes-Alnemri et al. (1994) found that overexpression of CPP32 in insect cells induced apoptosis. Coexpression of the 2 CPP32 subunits in insect cells also resulted in apoptosis. Tewari et al. (1995) showed that purified human ICE cleaved the Yama proenzyme into a proteolytically active form and that activated Yama cleaved PARP into the 85-kD apoptotic form.

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.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

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.

Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Rat, Mouse, By Heat
Western blot, 0.1-0.5μg/ml, Mouse, Rat

Validation Images & Assay Conditions

Click image to see more details

Figure 1. Western blot analysis of Caspase-3 (P17) using anti-Caspase-3 (P17) antibody (PA1961-1).
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 50ug of sample under reducing conditions.
Lane 1: Rat Heart Tissue Lysate
Lane 2: Rat Liver Tissue Lysate
Lane 3: Rat Thymus Tissue Lysate
Lane 4: Rat Spleen Tissue Lysate.
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-3 (P17) antigen affinity purified polyclonal antibody (Catalog # PA1961-1) 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 1:10000 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-3 (P17) at approximately 32KD,19KD. The expected band size for Caspase-3 (P17) is at 32KD,17KD.

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Figure 2. IHC analysis of Caspase-3 (P17)using anti-Caspase-3 (P17)antibody (PA1961-1).
Caspase-3 (P17)was detected in paraffin-embedded section of rat intestine tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Caspase-3 (P17)Antibody (PA1961-1) 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.

Click image to see more details

Figure 3. Western blot analysis of Caspase-3 (P17) using anti-Caspase-3 (P17) antibody (PA1961-1).
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 50ug of sample under reducing conditions.
Lane 1: mouse thymus tissue lysates,
Lane 2: mouse spleen tissue lysates,
Lane 3: mouse lung tissue lysates,
Lane 4: mouse brain tissue lysates,
Lane 5: mouse testis tissue 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-3 (P17) antigen affinity purified polyclonal antibody (Catalog # PA1961-1) 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 1:10000 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-3 (P17) at approximately 32KD,19KD. The expected band size for Caspase-3 (P17) is at 32KD,17KD.

Gene/Protein Information For CASP3 (Source: Uniprot.org, NCBI)

Alternative Names

Apopain; apoptosis-related cysteine protease; CASP3; CASP-3; caspase 3, apoptosis-related cysteine peptidase; Caspase3; Caspase-3; CPP32; CPP-32; CPP32B; CPP32SREBP cleavage activity 1; Cysteine protease CPP32; EC 3.4.22; EC 3.4.22.56; LICE-1; PARP cleavage protease; procaspase3; Protein Yama; SCA-1; YAMA CASP3 CPP32, CPP32B, SCA-1 caspase 3 caspase-3|CASP-3|CPP-32|PARP cleavage protease|SREBP cleavage activity 1|apopain|caspase 3, apoptosis-related cysteine peptidase|caspase 3, apoptosis-related cysteine protease|cysteine protease CPP32|procaspase3|protein Yama

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