ChIP assays were performed using NALM cells (a cell line derived from human pre-B leukemia), the anti-CIITA antibody (Cat. no. 49-1001), and optimized PCR primer sets. Each ChIP assay used sheared chromatin from 2 million cells and 5 µg (lane 3) and 7 µg (lane 4) of anti-CIITA antibody. HLA_DRA was used as positive PCR control, and HLA_DOB was used as negative PCR control. The ChIP results using the anti-CIITA antibody show a clear signal for the positive primer pair (HLA_DRA) with 5 µg and 7 µg of antibody per IP (lower panel, lanes 3 and 4) and show no signal for the negative primer pair (HLA_DOB) (upper panel, lanes 3 and 4). The input is shown in lane 1, and the anti-flag IP negative control antibody is shown in lane 2.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Raised against recombinant CIITA protein.|
|Contains||<0.1% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||5-7 ug|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
CIITA contains an acidic transcriptional activation domain, four LRRs (leucine-rich repeats) and a GTP binding domain. It is located in the nucleus and acts as a positive regulator of class II major histocompatibility complex gene transcription, and is referred to as the "master control factor" for the expression of these genes. CIITA also binds GTP and uses GTP binding to facilitate its own transport into the nucleus. Once in the nucleus it does not bind DNA but rather uses an intrinsic acetyltransferase (AT) activity to act in a coactivator-like fashion. Mutations in this gene have been associated with bare lymphocyte syndrome type II (also known as hereditary MHC class II deficiency or HLA class II-deficient combined immunodeficiency), increased susceptibility to rheumatoid arthritis, multiple sclerosis, and possibly myocardial infarction.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.