Cat. # | Size | Price | Inventory |
---|---|---|---|
48306S | 100 µl |
REACTIVITY | H |
SENSITIVITY | Endogenous |
MW (kDa) | |
Source/Isotype | Rabbit IgG |
Product Information
Application | Dilution |
---|---|
Immunofluorescence (Immunocytochemistry) | 1:12800 - 1:25600 |
Flow Cytometry (Fixed/Permeabilized) | 1:3200 - 1:12800 |
Achieve higher quality immunofluorescent images using the efficient and cost-effective, pre-made reagents in our #12727 Immunofluorescence Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Recommended Fluorochrome-conjugated Anti-Rabbit secondary antibodies:
NOTE: Cells should be grown, treated, fixed and stained directly in multi-well plates, chamber slides or on coverslips.
Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% formaldehyde diluted in 1X PBS.
NOTE: Formaldehyde is toxic, use only in a fume hood.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted November 2006
revised November 2013
Protocol Id: 24
All reagents required for this protocol may be efficiently purchased together in our Intracellular Flow Cytometry Kit (Methanol) #13593, or individually using the catalog numbers listed below.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: When including fluorescent cellular dyes in your experiment (including viability dyes, DNA dyes, etc.), please refer to the dye product page for the recommended protocol. Visit www.cellsignal.com for a full listing of cellular dyes validated for use in flow cytometry.
NOTE: Adherent cells or tissue should be dissociated and in single-cell suspension prior to fixation.
NOTE: Optimal centrifugation conditions will vary depending upon cell type and reagent volume. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells.
NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to fixation.
NOTE: Antibodies targeting CD markers or other extracellular proteins may be added prior to fixation if the epitope is disrupted by formaldehyde and/or methanol. The antibodies will remain bound to the target of interest during the fixation and permeabilization process. However, note that some fluorophores (including PE and APC) are damaged by methanol and thus should not be added prior to permeabilization. Conduct a small-scale experiment if you are unsure.
NOTE: Count cells using a hemocytometer or alternative method.
posted July 2009
revised June 2020
实验步骤编号:404
人
使用与人 BRD9 蛋白 Pro576 周围残基相对应的合成肽对动物进行免疫接种来产生单克隆抗体。
ATP 依赖性染色质重构复合体在调控各种胞核的过程中(例如:基因表达、DNA 复制和修复的过程中)起着重要作用 (1,2)。SWI/SNF 染色质重构复合体由 10 个以上的亚基和一个单分子的 ATP 酶催化性亚基 BRM 或 BRG1(仅两者之一)组成。这两种亚基的活性可促进组蛋白-DNA 接触的瓦解,组蛋白-DNA 接触能够导致染色质内部关键调控元件的可接近性发生改变 (2-5)。含有 SWI/SNF 复合体的 BRM/BRG1 被转录因子如核受体、p53、RB 和 BRCA1 召集至靶启动子以调控基因激活、细胞生长、细胞周期和分化过程 (1,6-9)。
GLTSCR1 及其横向同源物 GLTSCR1L 与 BRD9 一起被确定为非典型 BAF (ncBAF) 或 GBAF 复合体的独特亚基。该复合体包含 GLTSCR1 或 GLTSCR1L,而不是 ARID 亚基,同时也不包含 BAF45、BAF47 和 BAF57 亚基。GBAF 对应于不同于 PBAF 和典型 BAF 复合体的区域,并且被证实是 BAF 诱导癌症(如滑膜肉瘤和横纹肌样瘤)的一个合成致命靶标 (10-12)。
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