| Artikelnummer | GAS-002FOC |
|---|---|
| Artikelgruppe | Zellfixierung, Zellpermeabilisierung |
| Artikelbezeichnung | FIX & PERM Kit |
| Testprinzip | Erythrozytenlyse/Zellfixierung & -permeabilisierung, zur Färbung intrazellulärer Antigene für die Durchflusszytometrie |
| Inhalt | 1 x 5 ml Reagenz A (Fixierungsmedium), 1 x 5 ml Reagenz B (Permeabilisierungsmedium) |
| Zweckbestimmung | für In-vitro-Diagnostik |
| Packungsgröße | 15 Tests, Muster |
| Hersteller / Marke | Nordic-MUbio |
- GAS-002FOC – Datenblatt
- MSDS_FIX&PERM_EN-2019
ProductThis FIX&PERM® Cell Fixation and Permeabilization Kit contains 2 reagents: Fixation Medium (Reagent A) and Permeabilization Medium (Reagent B). It is intended for first fixing cells in suspension with Reagent A and then permeabilizing the cell membranes with Reagent B. This procedure gives antibodies access to intracellular structures and leaves the morphological scatter characteristics of cells intact. Specific formulations reduce background staining and allow simultaneous addition of permeabilization medium and fluorochrome labeled antibodies. FIX&PERM® is suitable for the analysis of normal and malignant leukocyte populations derived from various human biological samples (blood, bone marrow and others) using flow cytometry. Results must be put within the context of other diagnostic tests as well as the clinical history of the patient by a certified professional before final interpretation.Format: 4 x 5ml vial of Fixation Medium (Reagent A) and 4 x 5ml vial Permeabilization Medium (Reagent B). FIX&PERM® Reagents are designed for use with all commercially available flow cytometers. Alignment and compensation should be performed according to manufacturer´s instructions.The quality of each FIX&PERM® Lot is determined by fixation and permeabilization of well defined blood samples from representative donors and subsequent comparison of forward and side scatter characteristics of obtained leukocytes, as well as immunostaining efficiency for several membranous and cytoplasmic antigensFormulation: 4 x 5ml vial of Fixation Medium (Reagent A) and 4 x 5ml vial Permeabilization Medium (Reagent B). FIX&PERM Reagents are designed for use with all commercially available flow cytometers. Alignment and compensation should be performed according to manufacturer
ApplicationBiological fluids (blood, bone marrow, and others) must be collected under sterile conditions. Anticoagulation with EDTA or heparin is recommended. The samples should be stored at room temperature until used. For optimal results, samples should be processed and analyzed within 24 hours. Samples with high numbers of non-viable cells might cause false results, such cases require determination of cell viability with e.g. propidium iodide. All biological samples have to be handled with caution. Always consider them as potentially infective. Use appropriate precautions such as gloves, lab-coat, etc.
Fixation, permeabilization and staining procedure:
- For each sample to be analyzed add 50 µl of whole blood, bone marrow or mononuclear cell suspension in a 5ml tube
- Add 100 µl of Reagent A (Fixation Medium, stored and used at room temperature)
- Incubate for 15 minutes at room temperature
- Add 5ml phosphate buffered saline and centrifuge cells for 5 minutes at 300 g
- Remove supernatant and add to cell pellet 100 µl Reagent B (Permeabilization Medium) and 20 µl of the appropriate Nordic-MUbio monoclonal antibody conjugate
- Vortex at low speed for 1-2 seconds
- Incubate for 15 minutes at room temperature
- Wash cells with phosphate buffered saline as described above
- Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 ml 1.0 % formaldehyde and store them at 2-8°C in the dark. Analyze fixed cells within 24 hours.
Comments: Special cases (diluted bone marrow samples, other samples containing low soluble protein) might benefit from replenishment with plasma components before the FIX&PERM® treatment in order to create a milieu, which more closely resembles the stuation in anti-coagulated blood. For that pupose addition of IgG preparations (e.g. Beriglobulin P, ZLB Behring, final concentration 10mg/ml) and human serum albumin (e.g. human albumin "Behring" 20% – infusion solution, final concentration 40mg/ml) is recommended.
ReferencesRecent application paper:Nies KPH, Kraaijvanger R, Lindelauf KHK, Drent RJMR, Rutten RMJ, Ramaekers FCS, Leers MPG. Determination of the proliferative fractions in differentiating hematopoietic cell lineages of normal bone marrow. Cytometry A. 2018 Sep 3. doi: 10.1002/cyto.a.23564.
Here you can find more references and information about FIX&PERM.
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
抗体是属于у-球蛋白，或是淋巴细胞，都是对某种抗原反应而形成的；它附着在抗原上，起着抑制抗原的作用。如果抗体完全抑制了抗原，即使我们感染了病毒或细菌，也会在不知不觉中战胜疾病。相反，当抗原势力超过抗体时，疾病则会恶化。
一旦体内产生抗体时，当下次相同的病原体侵入时，会比前次更快发动免疫机构，将疾病防患于未然。许多病毒性疾病患者，经过一次治疗后便不再发病，是因为身体内已有了免疫的机构。
抗原与其所诱导产生的抗体或致敏淋巴细胞特异性结合的能力。抗原性的强弱与抗原分子的大小、化学成分、抗原决定簇的结构、抗原与被免疫动物亲缘关系的远近等有密切关系。通常认为抗原的分子量愈大、化学组成愈复杂、立体结构愈完整以及与被免疫动物的亲缘关系愈远，则抗原性愈强。抗原的物理状态也对抗原性发生影响，例如蛋白质，聚合状态的比单体的抗原性强，一般球形分子的比纤维形分子的抗原性强。抗原加入佐剂改变物理状态后，抗原性也得到增强。例如，分子量高达10万的明胶由于缺乏苯环氨基酸，稳定性较差，在进入机体后容易被酶降解成低分子物质，如果加入少量酪氨酸（苯环氨基酸），就能增强其抗原性。
中文名：抗原性
外文名：antigenicity
别称：免疫反应性
抗原性介绍：流感病毒按抗原性
1.抗原性是指抗原与其所诱导产生的抗体或致敏淋巴细胞特异性结合的能力。抗原性的强弱与抗原分子的大小、化学成分、抗原决定簇的结构、抗原与被免疫动物亲缘关系的远近等有密切关系。通常认为抗原的分子量愈大、化学组成愈复杂、立体结构愈完整以及与被免疫动物的亲缘关系愈远，则抗原性愈强。
2.免疫原性是指能够刺激机体形成特异抗体或致敏淋巴细胞的能力。即指抗原能刺激特定的免疫细胞，使免疫细胞活化、增殖、分化，最终产生免疫效应物质抗体和致敏淋巴细胞的特性。也指抗原刺激机体后，机体免疫系统能形成抗体或致敏T淋巴细胞的特异性免疫反应。
B细胞的抗原受体就是镶嵌在脂质双层中的膜结构蛋白，叫做膜表面免疫球蛋白（SmIg）。每个淋巴细胞克隆，只具有识别某一抗原决定簇的受体，每个淋巴细胞也只能产生某一种特异性抗体，这也是单克隆抗体只针对某一种抗原决定簇反应的基础。体内有千万种淋巴细胞克隆，所以机体能对各种不同的抗原性异物进行识别，从而发生免疫应答。
T细胞的抗原受体（T cell receptor, TCR）与CD3呈复合物形式存在于抗原特异性T细胞表面。在识别靶细胞过程中还有赖于非特异性的其他表面分子辅助，这些分子主要包括CD4、CD8、MHCI、MHCII类分子、淋巴细胞功能相关抗原-1（LFA-1）、细胞间黏附分子-1（ICAM-1，即CD54）、LFA-2（CD2）和 LFA-3（CD58）。TCR的多肽链是异质的，根据抗原结构和编码基因不同，已发现有a, b, g和d四种多肽链，据其组成不同，可分为两种类型，即abTCR和gdTCR。大多数TCR是abTCR，其中CD4+abTCR T细胞可识别非己MHCII类抗原（同种异体抗原）或自身MHCII类抗原与外来抗原复合物。abTCR识别外来抗原必须经过抗原提呈细胞（antigen presenting cell, APC）的加工处理，在外来抗原与APC表面MHC抗原结合成复合物才能被Th细胞识别。少数TCR由gd链组成，对gdTCR在体内的生理功能尚未完全清楚。
，表示体内感染了HBV，因而是一种特异性标志。HBsAg阳性见于：①急性乙型肝炎的潜伏期或急性期（大多短期阳性）；②HBV致的慢性肝病、迁延性和慢性活动性肝炎、肝炎后肝硬化或原发性肝癌等。③无症状携带者。
（2）抗HBs：表示曾感染过HBV，均已得到恢复，并且对HBV有一定的免疫力。
（3）HBcAg与抗HBc：由于
HBcAg主要存在于肝细胞核内，并仅存在于Dane颗粒中。因此，对病人血清不能检测HBcAg，而测抗HBc。血清内抗HBc阳性反映：①新近有过HBV感染；②体内有HBV增殖；③有助于诊断乙型肝炎。
（2）HBcAg和抗HBe：HBcAg的存在常表示病人血液有感染性。
HBcAg阳性揭示病人肝脏可能有慢性 损害，对预后判断有一定帮助。抗HBe阳性对病人可能有一定的保护力。
检测乙肝抗原与抗体的实际用途：
（1）筛选供血员。
（2）可作为乙肝病人或携带者的特异性诊断。
（3）对乙肝病人预后和转归提供参考。
（4）研究乙肝的流行病学，了解各地人群对乙肝的感染情况。
（5）判断人群对乙肝的免疫水平。
