Immunoprecipitation is a technique in which an antigen is isolated by binding to a specific antibody attached to a sedimentable matrix. It is also used to analyze protein fractions separated by other techniques such as gel filtration or density gradient sedimentation. The source of antigen for immunoprecipitation can be unlabeled cells or tissues, metabolically or intrinsically labeled cells, or in vitro-translated proteins. This unit describes a wide range of immunoprecipitation techniques, using either suspension or adherent cells lysed by various means.Usually, plasmas transfected cells or natural cells are ideal sample for immunoprecipitation.
A431 cells are a model cell line (epidermoid carcinoma) used in biomedical research. More specifically, they are used in studies of the cell cycle and cancer-associated cell signal pathways, because they express abnormally high levels of the Epidermal growth factor receptor (EGFR). As such they are often used as a positive control for EGFR expression. They contain no functional p53, a potent tumor suppressor gene,So they are highly sensitive to mitogenic stimuli. A431 cell line was established from an epidermoid carcinoma in the skin/epidermis of an 85- year-old female patient.
Daudi is a human B-lymphoblastoid cell line established from a Burkitt lymphoma of a 16 year old male patient. The cells express the receptor for the IgG-Fc fragment and complement receptors on their surface but do not express HLA antigens as surface markers.
THP1 is a human monocytic cell line isolated from an acute monocytic leukemia patient. Usually, it is used to test leukemia cell lines in immunocytochemical analysis of protein-protein interaction, and immunohistochemistry.
The Caco-2 cell line is a immortal cells of heterogeneous human epithelial colorectal adenocarcinoma cells, developed by the Sloan-Kettering Institute for Cancer Research through research conducted by Dr. Jorgen Fogh.
PC3 (PC-3) human prostate cancer cell line is one of the cell lines used in prostate cancer research. These cells are useful in investigating the biochemical changes in advanced prostatic cancer cells and in assessing their response to chemotherapeutic agents.
Raji cell line is the first continuous human cell line from hematopoietic origin. The cell line produce an unusual strain of Epstein-Barr virus and it will both transform cord blood lymphocytes and induce early antigens in Raji cells.
HL-60 cell line was derived from a 36-year-old woman with acute promyelocytic leukemia at the National Cancer Institute. HL-60 cells are predominantly a neutrophilic promyelocyte (precursor) which has been used for laboratory research on how certain kinds of blood cells are formed. The doubling time of HL-60 is about 36–48 hours which proliferates continuously in suspension culture in nutrient and antibiotic chemicals.
NIH 3T3 cells come from a cell line established in 1962 by two scientists at the Department of Pathology in the New York University School of Medicine, George Todaro and Howard Green. They originally obtained their 3T3 cells from Swiss albino mouse embryo tissue.The 3T3 cell line has become the standard fibroblast cell line. which refers to the cell transfer and inoculation protocol for the line, and means "3-day transfer, inoculum 3 x 10^5 cells." Using this protocol, the immortal cell line begins to thrive and stabilize in cell culture after about 20-30 generations of in vitro growth.
The A549 cell line was first developed in 1972 b D.J Giard, et al. through the removal and culturing of cancerous lung tissue in the explanted tumor of 58-year-old caucausian male. The cells produce adenocarcinomic alveolar basal epithelial cells with a modal chromosome number of 66. This type of cell squamous and performs the task of diffusing water, electrolytes and other substances across the surface of the alveoli in a normal lung.
K562 cell line is the first human immortalised myelogenous leukemia line to be established which is derived from a 53-year-old female CML patient in blast crisis. K562 cells are of the erythroleukemia type which are non-adherent and rounded, are positive for the bcr:abl fusion gene, and bear some proteomic resemblance to both undifferentiated granulocytes and erythrocytes.
RAW264.7 cells are macrophage-like cell line derived from Balb/c mice. They maintain a lot of properties of macrophages including NO production, phagocytosis (beads, other), extreme sensitivity to TLR agonists and motility. They are susceptible to genetic mutation, so freezer stocks must be made from early passage number cells.
SH-SY5Y is a human derived cell line used in scientific research. The original cell line, called SK-N-SH, from which it was subcloned was isolated from a bone marrow biopsy taken from a four-year-old female patient with neuroblastoma. SH-SY5Y cells are often used as in vitro research models of neuronal function and differentiation. They are adrenergic in phenotype but also express dopaminergic markers and, as such, have also been used to study Parkinson's Disease.
Jurkat cells are an immortalized cell line of T lymphocyte cells that are used to study acute T cell leukemia, T cell signaling, and the expression of various chemokine receptors susceptible to viral entry, particularly HIV. Jurkat cells are also useful in science because of their ability to secret interleukin 2. Their primary use, however, is to determine the mechanism of differential susceptibility of cancers to drugs and radiation. The Jurkat cell line (originally called JM) was established in the late 1970s from the peripheral blood of a 14 year old boy with T cell leukemia. Different derivatives of the Jurkat cell line can now be obtained from cell culture banks that have been mutated to lack certain genes.
HepG2 is a immortal cell line consisting of human liver carcinoma cells, isolated from the liver tissue of a 15-year-old Caucasian male who had a well-differentiated hepatocellular carcinoma. Hepatocellular carcinoma is, worldwide, the fifth most-common cancer.
MCF-7 cells was first isolated in 1970 from the breast tissue of a 69-year old Caucasian woman. Of the two mastectomies she received, the first revealed the removed tissue to be benign. Five years later, the second operation revealed malignant adenocarcinoma in a pleural effusion from which was taken cells for MCF-7. The woman was treated for breast cancer with radiotheraphy and hormonotherapy.
A HeLa cell /ˈhiːlɑː/, also Hela or hela cell, is a cell type in an immortal cell line used in scientific research. It is the oldest and most commonly used human cell line. The line was derived from cervical cancer cells taken on February 8, 1951, from Henrietta Lacks, a patient who eventually died of cervical cancer on October 4, 1951. The cell line was found to be remarkably durable and prolific — which has led to contamination of many other cell lines used in research.
Other positive cell lysate are:
molt-4 cell Lysate
SW480 cell Lysate
293T cell Lysate