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- Table of Contents
Facts about Autoimmune regulator.
Binds into nucleosomes (By similarity). Binds to chromatin and interacts selectively with histone H3 that is not methylated in'Lys-4', not phosphorylated in'Thr-3' rather than methylated in'Arg-2'.
Human | |
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Gene Name: | AIRE |
Uniprot: | O43918 |
Entrez: | 326 |
Belongs to: |
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No superfamily |
AIRE; APECED protein; APECED; APECEDAPSI; APS1; APSI; Autoimmune polyendocrinopathy candidiasis ectodermal dystrophy protein; autoimmune regulator (APECED protein)10APS1AIRE1; autoimmune regulator (autoimmune polyendocrinopathy candidiasis ectodermaldystrophy); Autoimmune Regulator; PGA1
Mass (kDA):
57.727 kDA
Human | |
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Location: | 21q22.3 |
Sequence: | 21; NC_000021.9 (44285876..44298648) |
Widely expressed. Expressed at higher level in thymus (medullary epithelial cells and monocyte-dendritic cells), pancreas, adrenal cortex and testis. Expressed at lower level in the spleen, fetal liver and lymph nodes. In secondary lymphoid organs, expressed in a discrete population of bone marrow-derived toleregenic antigen presenting cells (APCs) called extrathymic AIRE expressing cells (eTAC)(at protein level) (PubMed:23993652). Isoform 2 and isoform 3 seem to be less frequently expressed than isoform 1, if at all.
Nucleus. Cytoplasm. Predominantly nuclear but also cytoplasmic (PubMed:11274163, PubMed:14974083). Found in nuclear body-like structures (dots) and in a filamentous vimentin-like pattern (PubMed:11274163, PubMed:14974083, PubMed:26084028). Associated with tubular structures (PubMed:11274163, PubMed:14974083).
Boster Bio: How the AIRE Marker is used in research? We'll be discussing Autoradiography as well as Membrane Staining and how you can record your test's result using autoradiography film. We'll also discuss the efficacy of protein transfer using membrane staining. Finally, we'll cover the best applications for the AIRE Marker. Keep reading for more information.
Cell culture samples stained with the AIRE marker are a powerful tool for studying the integrity of cell membranes. It can be used to confirm that cells in aliquots have successfully transformed into monolayers. Furthermore, the ALI marker is able to detect smooth muscle cells. It is also useful to evaluate various experimental conditions. The AIRE marker can be used to identify proteins within various cell structures which include those in the immune system and blood.
The membrane is created during this step. The membrane is typically cut in preparation and shaped according to the gel size. The membrane should be handled with care because oil on the skin can interfere with the signal. To reduce background staining, wear gloves. The membrane is typically labeled in one corner. Side lanes are employed for pre-stained molecularweight markers. This stain can be used in conjunction with other immunodetection labels.
The AURE marker is part science and part art. Membrane staining requires a specific procedure. The method used and the type of bacteria used determine the time needed to stain a membrane. Always record the results in your lab notebook. Repeat the staining later in the semester, and try out different ones to determine which one produces the most effective results. If you're having difficulty identifying which antibodies are reactive with your samples, seek help from your instructor or lab friend.
AIRE-stained samples should be placed on the same slide as Gram-negative samples. This is to ensure that they stain the same way as Gram-negative samples. This helps prevent cell integrity problems. This process will allow you to distinguish bacteria from their DNA. And, once you have completed the process, you'll be able determine which Gram-positive bacteria and those that are negative.
Concentrates and pre-diluted RTU formats are readily available for the AIRE-specific antibody. Concentrates are easy to use and have lower costs at the beginning. They work with all staining systems that include manual and automated ones. They can be adjusted to suit the staining method and method used. The only disadvantage of using concentrates is the time required to prepare them.
Membrane staining is a vital method of determining the efficiency of protein transport. This staining technique uses staining of total proteins to identify the proteins that have been transferred from polyacrylamide gels into membranes. To detect proteins that have been removed from the gel, the membrane is stained with secondary antibodies. If the staining isn't consistent, it may indicate that the transfer process was not efficient. Uneven loading of the sample may be reflected in the intensities of the bands between lanes.
The membrane staining process used sera from healthy controls and human prostate cancer patients. These sera were transferred onto PVDF membranes. After transfer membranes were fixed by 50 percent methanol and 50 percent acetone. ECL Plus reagents were used to detect protein bands. After the immune response the sample was exposed to three different chemical treatments: ECL Plus and TBST.
The re-probed ability of PVDF and NC membranes was investigated. Different antibodies were employed to detect the bound proteins. The staining intensity of the proteins was assessed using densitometry and scanning. This method was used for clinical samples that have small sample sizes. Sometimes, antibodies are more efficient than lectins in detecting a variety of targets. The Boster Bio method is a great option for research and clinical use.
After the gel has been placed on the test plate the sample should be placed carefully to make sure there are no bubbles. The sample should be left to sit for between 30 to 60 minutes at 37°C. The gel should be solidified and polymerized at this point. If the gel has not formed yet, gently take it from the. Then mix the protein sample with 4X Dual Color Protein Loading Buffer, available from Boster. The sample should be added in the proper ratio of 3:1 to ensure most efficient results.
The results can be recorded using autoradiography film in the darkroom. The time for exposure varies on the effects of the developing solution. After the incubation time then the membrane should be rinsed three times with TBS Wash Buffer. The second antibody should be added. To get the best signal-to-noise ratio, the incubation time should be at minimum 10 minutes. Boster Bio is an excellent option for when the primary and second antibodies both transfer to the membrane.
You must use the latest fixer chemistry and developer chemistry in order to create high-quality autoradiographs. Use new fixer and developer chemistry every month to ensure that your autoradiographs are of the highest quality. Direct exposure will produce the highest resolution image, while fluorography reduces the sensitivity of the image and reduce its resolution. Typically, you will use a five-gallon developer tank and three trays. The film is transferred from one tray to the other using printing tongs as it is processed.
For the best results, you must use the appropriate safelight conditions. Choose a 15-W frosted or a Safelight Filter GBX-2. Do not use other safelights as they could create more background fog or cause an unintentional exposure of the image. The safelight should be placed at least four feet away from the film and allow it to develop for at least 30 seconds. Film will begin to fog in the majority of cases if it is exposed to the safelight for an extended period of time.
Autoradiography film has many advantages over digital imaging. Film captures medical images and is also available in digital formats. An autoradiography film is an emulsion made of radiation-sensitive crystals that are coated on a transparent base. After a few seconds of film exposure an image that is latent is formed. Then, a visible image is created. The image is processed by changing the image in the latent state to a visible one.
Depending on the technique used and the resolution desired the autoradiography film's sensitivity and resolution will vary. High-resolution autoradiography demands thin films with high radioisotope levels and long exposure durations. There are many methods that can calculate the exposure time for any level of radioactivity. The majority of autoradiography films are created in batches and radiomolecules are administered by various routes. In animal radioradiography radiomolecules can be administered via oral route, in the space between the ventricular walls, or in the peritoneum.
PMID: 9398839 by Nagamine K., et al. Positional cloning of the APECED gene.
PMID: 9398840 by Aaltonen J., et al. An autoimmune disease, APECED, caused by mutations in a novel gene featuring two PHD-type zinc-finger domains.