attribute (productName), human

Clone: REA1007
Clone xyz recognizes the human CD4 antigen, a 55 kDa single-pass type I membrane protein, also known as T4/Leu-3. CD4 is highly expressed on T helper cells and at a lower level on monocytes and dendritic cells. It is involved in the recognition of MHC class II/peptide complexes by the TCR heterodimers and is the receptor for the human immunodeficiency virus (HIV).
For 130-000-000, 130-000-001 the antibody titer is 1:50, for all other products use a titer of 1:11.

Applications

Reagent can be used for immunophenotyping by flow cytometry. Abnormal numbers of cells expressing this antigen or aberrant expression levels of the antigen can be expected in some disease states. It is important to understand the normal expression pattern for this antigen and its relationship to expression of other relevant antigens in order to perform appropriate analysis.
Expression of CD20 may be used as aid to diagnostic in the characterization of samples from individuals suspected with hematologic neoplasia.

Column capacity

Englischer Text, von dem es keine Übersetzung gibt.

Alternative names

ABC20, AD1

Background information

CD4 is an accessory molecule in the recognition of MHC class II/peptide complexes by the TCR heterodimers on CD4
+
T helper cells. CD4 is expressed on T helper cells and at a lower level on monocytes and dendritic cells. The CD4 molecule is the receptor for the human immunodeficiency virus. The CD4 antibody recognizes most thymocytes and about 65% of all peripheral blood T cells. See also
here
This is normal Text.

Detailed procedure

The isolation of exosomes or extracellular vesicles (EVs) is performed by positive selection using MicroBeads recognizing the tetraspanin proteins CD9, CD63, or CD81. First, EVs are magnetically labeled during a short incubation period. The labeled EVs are loaded onto a µ Column, which is placed in the magnetic field of a µMACS
Separator. The magnetically labeled EVs are retained within the column, while the unlabeled vesicels and cell components run through the column. After removing the column from the magnetic field, the intact EVs can either be collected by elution with Isolation Buffer, or directly lysed in the column and the protein in the lysates can be analyzed, e.g., by Western blotting.

Detailed separation procedure

One entire LRSC sample is labeled with StraightFrom™ LRSC CD14 MicroBeads and loaded onto LS Columns, which are placed in the magnetic field of a MultiMACS Cell24 Separator Plus or a MACS
®
Separator. After removing the column from the magnetic field, the magnetically retained CD14
+
cells can be eluted as the positively selected cell fraction.

Applications

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Downstream applications

Isolation or depletion of CD4
+
T helper cells is performed in many different fields of research, such as infectious diseases, autoimmune diseases, allergy, asthma, and tumor immunology. T helper cells isolated by MACS
®
Technology remain viable and functional. Therefore, they can be used for further functional studies, such as proliferation assays
10
and cytotoxicity assays
3
, as well as for the generation of T cell lines
9
. CD4
+
T cells purified with CD4 MicroBeads have also been cultured and analyzed for
in vitro
cytokine production.
4
Furthermore, they have been used in co‑culture experiments, e.g., for allogeneic stimulation by dendritic cells
1
or for B cell activation
2
.
T helper cells isolated by MACS
®
Technology were also used for studies on viral infections, e.g., for
in vitro
infections of cells with HIV
5
, to determine the infectivity of HIV-infected follicular dendritic cells
6
, immune reconstitution after anti-viral therapy
7
, and for monitoring of immune abnormalities in children of HIV-infected mothers
8
.

Columns

MS, LD, or autoMACS
®
Columns.

Quality description

Research-grade
cytokines are suitable for a wide variety of cell culture applications. They are sterile-filtered prior to lyophilization. Generally, endotoxin levels are <0.1 ng/μg (<1 EU/μg), and purities are >95%. The biological activity is tested in appropriate bioassays.

Biological activity

  • The specific activity is determined by proliferation assay according to Page
    et al.1
    using M07e cells. The proliferation assay was calibrated with the Non WHO Reference Material for human TPO (NIBSC code 03/124) provided by the National Institute for Biological Standards and Control.
  • premium grade: ≥ 2×
    10
    7
    U/mg
    (typical activity: ≥ 4×
    10
    8
    U/mg
    )
  • research grade: ≥ 1×
    10
    6
    U/mg
  • Specific activity: ≥ 3×
    10
    8
    U/mg