Caspase 3 p12 (Cleaved-Asp175) antibody

Catalog No. ABIN7251292

Product Details

Target: Caspase 3 p12

Binding Specificity: Cleaved-Asp175

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: Un-conjugated

Application: Western Blotting (WB), ELISA, Immunofluorescence (IF), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Characteristics: Polyclonal Antibody

Purification: Affinity purification

Immunogen: Synthesized peptide derived from the Internal region of human Caspase-3 p12

Isotype: IgG

Application Details

Application Notes: WB 1:500-2000, IHC 1:50-300, IF 1:50-300

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #104628 (Immunohistochemistry)

by: Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin

No.: #104628

Date: 03/15/2025

Antigen: Caspase 3 p12

Lot Number: DX05PXZ02179

Method validated: Immunohistochemistry

Positive Control:

The expression of caspase 3 at P5 was already attested in our laboratory (Lossi et al., 2004).

Negative Control:

A one control slice was processed for each experimental procedure, omitting the primary antibody.

Notes:

• Reference:

• Lossi L., Tamagno I. and Merighi A. (2004) “Molecular morphology of neuronal apoptosis: analysis of Caspase 3 activation during postnatal development of mouse cerebellar cortex.” J Mol Histol, 35(6):621-9.

Validation Images

Expression of cleaved caspase 3 (cCASP3) in P6 mouse cerebellum. Immunoreactive granule cells after immunostaining with ABIN7251291 and nuclear counterstaining with 4′,6-Diamidine-2′-phenylindole dihydrochloride (DAPI). EGL = external granular layer of the cerebellar cortex; IGL = internal granular layer of the cerebellar cortex.

Full Methods

Primary Antibody: ABIN7251291

Secondary Antibody: anti-rabbit Alexa Fluor 488 (Invitrogen. Lot 2541675)

Full Protocol:

• Sample analyzed: mouse brain at post-natal day (P) 5-6
• Paraffin-embedded slices (7 µm thick) were deparaffinized and rehydrated through a graded series of alcohols and distilled water (dH2O).
• Sections were blocked in ovalbumin 1% 1 hour at room temperature
• Antigen retrieval was performed in all the sections by microwave treatment (1 minute at 750W + 1 minute at 250W in Citrate buffer pH 6)
• 3x5 minutes washing in 0.01M PBS
• Sections were incubated with the primary antibody at the following dilutions of 1:50/1:100/1:200, overnight at room temperature (primary antibody was diluted in PLL/BSA/PBS diluent)
• 3x5 minutes washing in 0.01M PBS
• Incubation with the anti-rabbit secondary antibody 1:400 in PLL/BSA/PBS diluent, 1 hour at room temperature.
• 3x5 minutes washing in 0.01M PBS
• Nuclear counterstaining was performed with DAPI 1µg/mL, 2 minutes
• 2x5 minutes washing in 0.01M PBS
• Specimens were then mounted in Fluoroshield mounting medium (Sigma)

Experimental Notes:

The antibody works in IHC-P at concentrations from 1:50-1:200.

Handling

Format: Liquid

Concentration: 1 mg/mL

Buffer: PBS with 0.02 % sodium azide, 0.5 % BSA and 50 % glycerol, pH 7.4

Preservative: Sodium azide

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

Storage: -20 °C

Storage Comment: Store at -20°C. Avoid freeze / thaw cycles.

Target Details

Target: Caspase 3 p12

Alternative Name: CASP3 p12

Background: Involved in the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp-Gly-217' bond. Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9. Involved in the cleavage of huntingtin.

Molecular Weight:

Observed_MW: 20+35 kDa

Calculated_MW: 32 kDa

UniProt: P42574