Genox

Anti-7-Ketocholesterol Monoclonal Antibody
(Anti 7-KC monoclonal antibody, clone #35A)
(Catalog #MKC-100 for 100 microgram vial; MKC-020 for 20 microgram vial)

Product Insert (pdf)pdf

What is 7-ketocholesterol?
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.

mono2a

 

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 1mg/mL
(please take care that only a small volume is in the vial).

Specificity

:

Specific for 7KC

Class

:

Mouse IgG

Storage

:

Less than -20°C

Application

:

Immunohistochemistry?It is recommended that the
antibody be tried at 5-10 micro g/mL on frozen tissue.
Not applicable to paraffin embedded tissues.

Specificity of anti 7-KC antibody.

 


Immuno-histochemical detection of 7KC in atherosclerotic aorta.
Noriyuki Shibata et al. Tokyo Women’s Medical University

 

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%
non-immune animal serum in PBS.

6)

Block endogenous 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-immuno-peroxidase
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.

The ‘Anti-MKC Monoclonal Antibody’ is manufactured in Japan
by the Life Science Products Division of the Nippon Oils and Fats
Corporation, Japan. Genox is the distributor of this product
for users in North and South America.

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Note: Genox’s Products are for RESEARCH USE ONLY.
Users of Genox’s products are strongly advised NOT TO USE THEM FOR CLINICAL/DIAGNOSTIC APPLICATIONS. 

The provision of test reports, generated by using Genox's products, to individuals
or to treating physicians for the diagnosis, prevention, treatment and control of any human disease or impairment of, or the assessment of the health, nutritional, or medical condition of individuals is expressly prohibited by law.
  42 U.S.C. § 263a (2006); 42 C.F.R. § 493.3 (2006); COMAR 10.10.01.02 (2006). Genox is not a CLIA certified laboratory.

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