Caspase 3 ELISA Kit

Catalog No. ABIN4886354

The Human and Mouse Caspase 3 ELISA Kit (ABIN4886354) is a Colorimetric ELISA Kit designed to quantify Human and Mouse Caspase 3.

Quick Overview for Caspase 3 ELISA Kit (ABIN4886354)

Target: Caspase 3 (CASP3)

Binding Specificity: Cleaved-Asp175

Reactivity: Human, Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details Caspase 3 ELISA Kit

Purpose: CASP-3 (D175) ELISA Kit. This ELISA is for measuring cleaved CASP-3 (Asp-175) in human cell lysates.

Analytical Method: Semi-Quantitative

Specificity: The antibody pair provided in this kit recognizes human/mouse cleaved-caspase-3 cleaved at site Aspartic Acid-175

Characteristics:

• Rapidly measures cleaved protein in lysates
• Screen numerous different cell lysates without performing a Western Blot analysis
• Minimal hands-on time, convenient, and non-radioactive material

Components:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Anti-cleaved Antibody
• HRP-Conjugated Secondary Antibody
• Assay Diluent
• TMB One-Step Substrate
• Stop Solution
• Lysis Buffer
• Positive Control Sample

,

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Anti-cleaved Antibody
• HRP-Conjugated Secondary Antibody
• Assay Diluent
• TMB One-Step Substrate
• Stop Solution
• Lysis Buffer
• Positive Control Sample

Material not included:

• Distilled or deionized water
• 100 mL and 1 liter graduated cylinders
• Tubes to prepare sample dilutions
• Protease and Phosphatase inhibitors
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• Benchtop rocker or shaker
• Microplate reader capable of measuring absorbance at 450 nm

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Plate: Pre-coated

Protocol:

1. Prepare all reagents and samples as instructed in the manual.
2. Add 100 μL of sample or positive control to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared primary antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared 1X HRP-Streptavidin to each well.
7. Incubate 1 h at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Restrictions: For Research Use only

Handling

Storage: -20 °C

Storage Comment: Upon receipt, the kit should be stored at -20°C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4°C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge, and store at -20°C. Item D, store at 2-8°C for up to one month (store at -20°C for up to 6 months, avoid repeated freeze-thaw cycles). Reconstituted Positive Control (Item K) should be stored at -70°C.

Expiry Date: 6 months

Target Details for Caspase 3

Target: Caspase 3 (CASP3)

Alternative Name: CASP3

Background:

Gene Symbols: CASP3, CPP32

Background: Cleaved-Caspase-3 (Asp175) ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human and mouse cell lysates. By determining cleaved Caspase-3 protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.

This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human and mouse Cleaved-Caspase-3. An anti-Caspase-3 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and Caspase-3 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti-Cleaved-Caspase-3 (Asp175) antibody is used to detect Cleaved Caspase-3. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Cleaved Caspase-3 (Asp175) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Protein Name (Synonyms): Caspase-3 (CASP-3) (EC 3.4.22.56) (Apopain) (Cysteine protease CPP32) (CPP-32) (Protein Yama) (SREBP cleavage activity 1) (SCA-1) [Cleaved into: Caspase-3 subunit p17 Caspase-3 subunit p12]

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