Anti 7-ketocholesterol antibody
(Anti 7-KC monoclonal antibody, clone #35A)
FOR RESEARCH USE ONLY
About 7-ketocholesterol (7-KC)
Cholesterol oxidation products, especially
7-Ketocholesterol (7KC), have been the focus of much attention because they are present in human atherosclerotic plaque and display
a wide range of atherogenic properties in vitro and to some extant in vivo. This antibody is specific for 7KC. This antibody is useful to
detect 7KC in tissue samples. A frozen tissue sample is recommended for immunohistochemistry.
Specifications
Clone #:
35A
Antigen:
7KC-modified keyhole-lympet hemocyanine.
Form:
Protein A purified. Frozen in 10mM PBS containing 0.1% NaN3 and 0.5% BSA.
IgG concentration is 1 mg/mL (please take care that only a small volume is in the vial).
Specificity:
Specific for 7KC
Class:
Mouse IgG1 (kappa)
Application:
Immunohistochemistryˇ¨
It is recommended that the antibody be tried at 5-10 µg/mL on frozen tissue.
Not applicable to paraffin embedded tissues.
Product name
Code
Content
Anti 7-ketocholesterol (7-KC) monoclonal antibody
MKC-020n
20 µg IgG
MKC-100n
100 µg IgG
FOR RESEARCH USE ONLY. Not for diagnostic, medical or other use.
Immunohistochemistry
Procedure
1)
Samples were fixed in 10% formalin, immersed in 30% sucrose in PBS,
embedded at OCT (Sakura, Tokyo, Japan), and stored at 80 degree C.
2)
Prepare 3-7um thick sections from OCT compound embedded fresh
frozen samples.
3)
Wash OCT compound with PBS (pH7.6).
Please take care not to wash with 100% acetone.
4)
Frozen sections were quenched for several minutes with
1% EDTA, 3% hydrogen peroxide /PBS.
5)
Sections are rinsed in PBS, and pretreated for 30 min
with 3% nonimmune animal serum in PBS.
6)
Block endgenous avidin biotin activity.
7)
Dilute anti 7-KC antibody with 1% BSA, 0.1% NaN3
, 15mM EDTA in PBS (pH7.6).
8)
Sections were incubated overnight at 4 degree C with
primary antibody at a concentration of 10 ug/mL.
9)
Place the sections at room temperature, and wash with PBS
for 5 minutes x 3 times.
10)
Antibody binding was visualized by the avidin-biotin-immunoperoxidase
complex method using the Vectastain ABC kit (Vector, Burlingame, CA) according to the manufacturer's instructions.
11)
Use 3,3-Diaminobenzidine tetrahydrochloride (DAB) as the chromogen.
12)
Counterstain by Hematoxylin or Eosin.
Technical Note
1)
Frozen materials only.
2)
Sections from which the primary antibodies were omitted served
as negative reaction controls.
Positive control
Aortic wall of atherosclerosis.
FOR RESEARCH USE ONLY. Not for diagnostic, medical or other use.
