Caspase 3 antibody

Catalog No. ABIN3188045

This anti-Caspase 3 antibody is a Mouse Monoclonal antibody detecting Caspase 3 in WB and IHC. Suitable for Human, Mouse and Rat. Independently validated for use in Western Blotting. This Primary Antibody has been cited in 1 publication.

Quick Overview for Caspase 3 antibody (ABIN3188045)

Target: Caspase 3 (CASP3)

Reactivity: Human, Mouse, Rat

Host: Mouse

Clonality: Monoclonal

Conjugate: This Caspase 3 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC)

Clone: 5E1

Product Details anti-Caspase 3 Antibody

Specificity: The antibody detects endogenous Active Caspase-3 protein.

Characteristics: Mouse Monoclonal to Active Caspase-3.

Purification: The antibody was affinity-purified from mouse ascites by affinity-chromatography using epitope-specific immunogen.

Immunogen: Recombinant Protein

Isotype: IgG1

Application Details

Application Notes: WB 1:500-1000, IHC 1:100-200

Restrictions: For Research Use only

Independent Validation (WB)

Validation #100061 (Western Blotting)

by: Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin

No.: #100061

Date: 08/31/2016

Antigen: Caspase 3 (active)

Method validated: Western Blotting

Positive Control: Mouse cerebellar tissue samples at post natal day P5

Notes: The antibody recognized both the active and non-active form of Caspase3 and worked well in the experimental conditions tested. The product is therefore indicated for the WB use.

Validation Images

P5 mouse cerebellum, 30 seconds exposition. 60µg (left) and 30µg (right) protein loaded. The inactive procaspase 3 is visible at a MW of approximately 32kDa. The p17 subunit of active Casp3 is revealed between the 15kDa and 20kDa bands of the MW marker.

Full Methods

Secondary Antibody: ABIN3179097

Full Protocol:

• Mouse cerebellar tissue samples at post natal day (P) 5 were lysed in 150 µL of lysis buffer.
• Protein concentration was determined using Bradford reagen (Sigma B6916) according to the manufacturer's recommendation.
• 30 and 60µg total protein were denatured in sample buffer for 5min at 95°C and then cooled on ice.
• Proteins were separated on a denaturing homemade 15% SDS-PAGE gel at a constant voltage of 180V for 70min.
• Proteins were transferred in a transfer buffer to nitrocellulose membrane (Amersham Biosciences, RPN203D, lot 18100/66/0207) at constant amperage of 400mA for 60min.
• Blocking of the membrane with blocking solution (0.01M PBS, 0.1% tween, 2% Bovine Serum Albumin) for 1h at RT.
• Incubation with the primary antibody ABIN3179097 diluted 1:1000 in blocking solution ON at 4°C.
• 3 washes in PBS-0.1% tween for 15min at RT.
• Incubation with the anti-mouse horseradish peroxidase (HRP) secondary antibody (Santa Cruz, SC2005, lot H0406) diluted 1:10000 in blocking solution for 1h at RT.
• Washes in PBS-0.1% tween for 15min at RT.
• Chemiluminescence detection using homemade ECL (1min incubation at RT) and revealed on HYPERFILM EGL (Amersham Biosciences, RPN2013K, lot 04450300115); exposure 30sec.
• Proteins were revealed using the ECL technique on western blot films for 30 seconds.

Experimental Notes:

• A band around 30-35 kDa and a band around 17kDa were detected with the experimental procedures previously described.
• A protein load of 30µg is recommended, as in the 60µg load, weak non-specific bands were detected around 15 and 30kDa.

Handling

Format: Liquid

Buffer: Liquid in PBS containing 50 % glycerol, 0.5 % BSA and 0.02 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeated freeze/thaw cycles.

Storage: -20 °C

Storage Comment: Store at -20°C.

References for anti-Caspase 3 antibody (ABIN3188045)

Ding, Loganathan, Zhou, Sayour, Brlecic, Radovits, Domain, Korkmaz, Karck, Szabó, Korkmaz-Icöz: "Alpha-1-Antitrypsin Protects Vascular Grafts of Brain-Dead Rats Against Ischemia/Reperfusion Injury." in: The Journal of surgical research, Vol. 283, pp. 953-964, (2023) (PubMed).

Target Details for Caspase 3

Target: Caspase 3 (CASP3)

Alternative Name: Caspase-3

Gene ID: 836

UniProt: P42574

Pathways: Apoptosis, Caspase Cascade in Apoptosis, Sensory Perception of Sound, ER-Nucleus Signaling, Positive Regulation of Endopeptidase Activity, Activated T Cell Proliferation