Caspase 9 Activity Assay Kit (by Spectrophotometry)[313]

    <
    >

      Catalog number:E-CK-A313

      Size:
      • 20 Assays
      • 50 Assays
      • 100 Assays
      Qty:
      - +
      Price: $68

      Lead Time: Order now, ship in 3 daysWelcome to order from local distributors.

      Add to cart Compare Bulk request Manual

      Introduction

      Elabscience® Caspase 9 Activity Assay Kit can detect the activity of Caspase 9 in cells or tissue lysis.


      Detection principle

      Caspase (Cysteine-requiring Aspartate Protease) is a protease family that plays an important role in the process of apoptosis. Caspase 9 (also known as ICE-LAP6, or mch6) is considered to be a caspase in the upstream of apoptotic signal transduction. When mitochondria release cytochrome c, Caspase 9 can form complex with cytochrome c and Apaf1, and be activated at the same time. Activated Caspase 9 can activate Caspase 3 which is the key enzyme of cell apoptosis, thereby promoting subsequent apoptotic signals. Caspase 9 exists in the form of prozyme in the normal state and has no activity. However, Caspase 9 is activated in the apoptotic stage and participates in the apoptotic process.

      This kit is used to conjugate Caspase 9 sequence-specific peptides acetyl-Leu-Glu-His-Asp p-nitroanilide (Ac-LEHD-pNA) to yellow group p-nitroaniline (pNA). When the substrate is cut by Caspase 9, the yellow group pNA is dissociated. pNA has an absorption peak at 405 nm. Measure the OD value at 405 nm and then Caspase 9 activity can be calculated accordingly.


      Componenta


      Cat.

      Products

      20 Assays

      50 Assays

      100 Assays

      Storage

      E-CK-A313A

            Lysis Buffer

      3.0 mL

      7.5 mL

      15 mL

      -20°C

      E-CK-A313B

            2 ×Reaction Buffer

      1.0 mL

      1.25 mLx 2

      5.0 mL

      -20°C

      E-CK-A313C

            Ac-LEHD-pNA

      100 μL

      250 μL

      500 μL

      -20°C

      E-CK-A313D

            DTT

      50 μL

      100 μL

      150 μL

      -20°C

      Manual

       One Copy


      Reagents not included

      PBS, Protein Quantitative Kit (Bradford, optional).


      Storage

      Store at -20°C. Ac-LEHD-pNA[E-CK-A313C] should be stored in dark. Avoid freeze / thaw cycles.

       

      Note

      1.  For maximal assay performance, this reagent should be used within 12 months. Avoid freeze / thaw cycles.

      2.  There is reducing agent (DTT) in the Lysis Buffer,It is recommended to Elabscience® Total Protein (TP) Colorimetric Assay Kit (Coomassie Brilliant Blue Method) (It is recommended to use Elabscience® E-BC-K168-S) to measure the protein concentration instead of BCA method.

      3.  It has been reported that the activity of Caspase 9 can’t be detected in few types of apoptosis, which may be due to the existence of a mechanism independent of the activation of Caspase 9, and other signaling pathways in the mechanism of apoptosis need to be considered. In this case, there is no significant change in the activity of Caspase 9 by using this kit.

      4.  Lysis Buffer in this kit can be used in other Caspase activity assay kits by Elabscience®. And the protein sample can test other Caspase activity.

      5.  pNA (4-nitroaniline) is toxic. Please be careful when operating, and pay attention to effective protection to avoid direct contact with human body or inhalation. pNA solidifies at lower temperatures and sticks to the bottom, wall or cap of centrifugal tube. It can be incubated in water bath at 20~25°C for a short time until it is completely melted.

      6.  When the activity of Caspase 9 in the sample is very low. Confirm whether the phenomenon of apoptosis is obvious or not firstly. If the apoptosis is obvious and we confirm that Caspase can be activated, please adjust the time of apoptosis and find a time point which Caspase 9 activation is stronger. And then repeat the test.

      7.  This kit is for research use only. For your safety and health, please wear lab clothes and gloves. Instructions should be followed strictly, changes of operation may result in unreliable results.

      submit review
      • Show all (0)
      • Reviews (0)
      • Q&A (0)
      ... Show All Show Less
      MSDS for ELISA ISO 9001

      Browsing History


        People Also Bought

        Apply for
        *Product Name:
        *Catalog Number:
        *Name:
        *Email:
        *Message:
        *Country:
        How do you know
        Elabscience:

        *Captcha: