Q: Could I use HeLa cell lysate as a positive control for Caspase 9 primary antibodies in WB?

A: After looking through our testing it does appear you should be fine using HeLa as a positive control cell lysate for Caspase 9 in Western Blot, as we have used this sample successfully in our QC tests and shown expression of Caspase 9.

Human Caspase-9 ELISA Kit (Colorimetric)

Novus Biologicals | Catalog # NBP1-83734

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Product Specifications
Precision
Recovery
Scientific Data
Kit Contents
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas
Related Blogs

Key Product Details

Sample Type & Volume Required Per Well

Cell culture supernatant, cell lysate, serum (50 uL)

Sensitivity

0.4 ng/mL

Assay Range

1.6 - 100 ng/mL

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Product Specifications

Assay Type

Sandwich ELISA

Kit Type

ELISA Kit (Colorimetric)

Species

Human

Specificity

The interference of circulating factors of the immune system was evaluated by spiking these proteins at physiologically relevant concentrations into a human Caspase 9 positive sample. No crossreactivity was detected.

Description

The human Caspase-9 ELISA is an enzyme-linked immunosorbent assay for quantitative detection.

Precision

Intra-Assay Precision (Precision within an assay) CV% 6.6%

Inter-Assay Precision (Precision between assays) CV% 9.0%

Recovery for Human Caspase-9 ELISA Kit (Colorimetric)

Recovery

The amount of endogenous human Caspase-9 in unspiked lysates was subtracted from the spike values. The overall mean recovery was 103%.

Scientific Data Images for Human Caspase-9 ELISA Kit (Colorimetric)

ELISA: Human Caspase-9 ELISA Kit (Colorimetric) [NBP1-83734]

ELISA: Caspase-9 ELISA Kit [NBP1-83734] - Standard curve

Kit Contents for Human Caspase-9 ELISA Kit (Colorimetric)

Adhesive Films
Aluminium pouch(es) with a Microwell Plate coated with monoclonal antibody to human Caspase-9
Anti-human Caspase-9 polyclonal Detection Antibody (rabbit)
Anti-rabbit-IgG-HRP
Assay Buffer Concentrate 20x (PBS with 1% Tween 20 and 10% BSA)
Human Caspase-9 Standard lyophilized, 200 ng/ml upon reconstitution
Lysis Buffer 10x
Sample Diluent
Stop Solution (1M Phosphoric acid)
Substrate Solution (tetramethyl-benzidine)
Wash Buffer Concentrate 20x (PBS with 1% Tween 20)

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Storage of components varies. See protocol for specific instructions.

Background: Caspase-9

Caspases are the executioners of apoptosis. This cysteine protease family consists of more than 10 related members characterized by almost absolute specificity for aspartic acid in the P1 position. Caspases are synthesized as inactive proenzymes comprising an N-terminal peptide together with one large and one small subunit. Activation of caspases during apoptosis results in the cleavage of critical cellular substrates so precipitating the dramatic morphological changes of apoptosis. Caspase-9, formerly called Apaf-3, binds to Apaf-1 in the presence of cytochrome c and dATP (3), which leads to caspase-9 activation. Activated caspase-9 in turn cleaves and activates caspase-3; caspase-9 thus is the most upstream member of the apoptotic protease cascade that is triggered by cytochrome c and dATP. Caspase-9 is essentially localized in mitochondria, the disruption of the outer mitochondrial membrane occurring early during apoptosis is critical for its subcellular redistribution and activation. Apart from cleavage of caspase-3, the downstream enzymes caspase-6 and 7 are targets of caspase-9 activity. Caspase-9 forms dimers in the course of activation. Cytochrome c/Apaf-1/caspase-9 form a so called apoptosome, amplifying the caspase cascade.

Alternate Names

CASP9, Caspase9

Gene Symbol

CASP9

Additional Caspase-9 Products

Product Documents for Human Caspase-9 ELISA Kit (Colorimetric)

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Download Manual - Human Caspase-9 ELISA Kit (Colorimetric)

Certificate of Analysis

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Product Specific Notices for Human Caspase-9 ELISA Kit (Colorimetric)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. ELISA Kits are guaranteed for 6 months from date of receipt.

Related Research Areas

Cysteine Proteases and Regulators

Citations for Human Caspase-9 ELISA Kit (Colorimetric)

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