Name: C7 Depleted Serum
SKU: A324C
Price: 100.00 USD
Availability: InStock

Description

General Description

Normal human serum depleted of complement C7 protein by immunoaffinity chromatography. The product is tested for the absence of C7 by functional assays for classical pathway activity and alternative pathway activity and for C7 protein by double immunodiffusion. C7-Dpl is certified to possess functional classical and alternative pathways for complement activation up to the step of C7 binding to C5b,6 (Law, S.K.A. and Reid, K.B.M. (1995); Morgan, B.P. ed. (2000); Dodds, A.W. and Sim, R.B. editors (1997)). Functional complement systems can be reconstituted by addition of purified C7 protein (56 µg/mL) indicating that all other complement components necessary for complement activation are present.

Specifications

Sizes Available: 1.0 mL/vial
Concentration: >50 mg protein/mL (see Certificate of Analysis for actual conc.)
Form: Frozen liquid
Activity: >80% versus normal human serum standard
Purity: No C7 detectable by immunodiffusion
Buffer: 10 mM sodium phosphate, 145 mM NaCl, pH 7.3
Preservative: None, 0.22 µm filtered
Storage: -70o C or below. Minimize freeze/thaw cycles.
Source: Normal human serum (shown by certified tests to be negative for HBsAg and for antibodies to HCV, HIV-1 and HIV-II).
Precautions: Use normal precautions for handling human blood products.
Origin: Manufactured in the USA.

Safety and Handling

Precautions/Toxicity/Hazards

The source is human serum, therefore precautions appropriate for handling any blood-derived product must be used even though the source was shown by certified tests to be negative for HBsAg and for antibodies to HCV, HIV-1 and HIV-II.
Hazard Code: B WGK Germany 3
MSDS is available upon request.

Additional Information

Physical Characteristics

Clear, straw-colored liquid containing all proteins of normal human serum except complement component C7.

Function

The depleted serum is tested for remaining classical pathway hemolytic activity using antibody-sensitized sheep erythrocytes (CompTech #B200) and for alternative pathway function using rabbit erythrocytes (CompTech #B300). The depleted serum is
reconstituted with 56 µg/mL C7 (CompTech #A124) and retested to verify that a hemolytic function is restored. The Certificate of Analysis provided with each lot gives a description of the assays and specific titers for the depleted and reconstituted sera compared to normal human serum.

Assays

The unit of classical pathway activity is the CH50. A similar unit, the C7H50, is used to quantitate the activity of C7 and C7-Dpl. A C7H50 unit is the amount of C7 needed to lyse 50% of 3 x 107 EA cells (antibody- sensitized sheep erythrocytes (CompTech #B200)) when that amount of C7 (CompTech #A124) is incubated with the recommended volume of C7-Dpl in GVB++ in a total volume of 500 µL for 30 min at 37o C. This amount of C7 indicates the sensitivity of the assay for C7 which is typically about <5 ng C7 with 25 µL C7-Dpl. See the Certificate of Analysis for lot specific values. Controls without C7 exhibit typically <5 % lysis. After full reconstitution (56 µg C7/mL C7-Dpl) lysis should be 100% in this assay.

Applications

C7-Dpl is used to assay C7 hemolytic activity in samples and to supply an activating system that is incapable of activating the membrane attack complex (C5b-9) of complement past the C5b,6 stage.