Anti-Caspase-3(P17)/CASP3 Antibody Picoband®

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.

Mean gray value analyzed according to western blotting results of IL-6, caspase-3, and cleaved-caspase-3.
Index in PubMed under a CC BY license. PMID: 34975719

Experimental workflow. One hundred four rats were randomly divided into five groups: group S (sham, n = 20), group M (middle cerebral artery occlusion [MCAO], n = 28), group H2M (intermittent hypobaric hypoxia preconditioned MCAO group, 2 h/day, n = 20), group H6M (intermittent hypobaric hypoxia preconditioned MCAO group, 6 h/day, n = 28), and group HpM (persistent hypobaric hypoxia preconditioned MCAO group, n = 28). Behavioral tests and morphological staining (TTC staining) were used to analyze the severity of infarction. Total protein expression of NeuN (a specific marker of mature neurons), caspase-3, cleaved-caspase-3, and IL-6 was estimated using western blotting, which explained the severity of injury from different perspectives. Ultrastructural changes were observed under a transmission electron microscope. The most effective pretreatment group was selected for further label-free proteomic study and provided a reliable direction for mechanism exploration. Western blotting was used to verify the expression of the target protein, and key markers for the biological process were detected using immunofluorescence. caspase-3, cysteinyl aspartate specific proteinase 3; IL-6, interleukin 6; NeuN, neuron-specific nuclear protein; TTC, 2,3,5-triphenyl tetrazolium chloride.
Index in PubMed under a CC BY license. PMID: 34975719

Relative expression of IL-6, caspase-3, and cleaved-caspase-3. (A) Immunoblot results of IL-6, caspase-3, and cleaved-caspase-3. (B) Analysis of IL-6 relative expression. One-way ANOVA showed differences among the five groups, F = 10.86, p < 0.0001. (C) Analysis of caspase-3 relative expression. One-way ANOVA showed differences among the five groups, F = 8.50, p = 0.0004. (D) Analysis of cleaved-caspase-3 relative expression. One-way ANOVA showed differences among the five groups, F = 17.36, p < 0.0001. ANOVA, analysis of variance; caspase-3, cysteinyl aspartate specific proteinase 3; IL-6, interleukin 6.
Index in PubMed under a CC BY license. PMID: 34975719

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.

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.