NTAL (6A454)
英文名称: NTAL (6A454)
型号:null    抗体货号: sc-71731
价格:请致电:010-57128832,18610462672
品牌: 美国原装    产品商标: santa cruz biotechno

现货1750促销

 

BACKGROUND
Non-T cell activation linker (NTAL), a transmembrane adaptor protein, is
also designated membrane-associated adapter molecule, Williams-Beuren
syndrome chromosome region 15 protein or LAB (linker of activated B cells).
NTAL is present in membrane microdomains (rafts) of B cells, NK cells and
myeloid cells, and in monocytes and mast cells, but not in resting T lymphocytes.
NTAL becomes rapidly tyrosine-phosphorylated upon cross-linking of
the B cell receptor (BCR) or of high-affinity Fc γ and Fc ε receptors of myeloid
cells and then associates with the cytoplasmic signaling molecules. NTAL is
highly expressed in spleen, lymph node germinal centers and peripheral blood
lymphocytes. Defects in the gene encoding for NTAL may cause the musculoskeletal
and cardiovascular abnormalities that characterize the rare developmental
disorder Williams-Beuren syndrome (WBS).

REFERENCES
1. Martindale, D.W., Wilson, M.D., Wang, D., Burke, R.D., Chen, X., Duronio,
V. and Koop, B.F. 2000. Comparative genomic sequence analysis of the
Williams syndrome region (LIMK1-RFC2) of human chromosome 7q11.23.
Mamm. Genome 11: 890-898.
2. Janssen, E., Zhu, M., Zhang, W., Koonpaew, S. and Zhang, W. 2003. LAB:
a new membrane-associated adaptor molecule in B cell activation. Nat.
Immunol. 4: 117-123.
3. Tkaczyk, C., Horejsi, V., Iwaki, S., Draber, P., Samelson, L.E., Satterthwaite,
A.B., Nahm, D.H., Metcalfe, D.D. and Gilfillan, A.M. 2004. NTAL phosphorylation
is a pivotal link between the signaling cascades leading to human
mast cell degranulation following Kit activation and Fc ε RI aggregation.
Blood 104: 207-214.
4. Koonpaew, S., Janssen, E., Zhu, M. and Zhang, W. 2004. The importance
of three membrane-distal tyrosines in the adaptor protein NTAL/LAB. J.
Biol. Chem. 279: 11229-11235.
5. Tedoldi, S., Paterson, J.C., Hansmann, M.L., Natkunam, Y., Rüdiger, T.,
Angelisova, P., Du, M.Q., Roberton, H., Roncador, G., Sanchez, L., Pozzobon,
M., Masir, N., Barry, R., Pileri, S., Mason, D.Y., Marafioti, T. and Horejsí, V.
2005. Transmembrane adaptor molecules: a new category of lymphoid-cell
markers. Blood 107: 213-221.

 

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      M-MLV 逆转录酶 ,Rnase H-,点突变

M-MLV逆转录酶是一种依赖于RNA和DNA的DNA聚合酶。它可以单链RNA、DNA或RNA-DNA杂合链为模板,以RNA或DNA为引物启动DNA合成。

该酶带有针对RNA-DNA杂合链中RNA的RNase H活性。

本产品来源于带有编码莫洛尼氏鼠白血病病毒(M-MLV)逆转录酶的pol基因片段的E.coli菌株。

 

特点:

· 产量更高,第一链cDNA可达9kb;

· 37时活性最高;

· 可用于掺入修饰过的核苷酸(e.g. Cy3-, Cy5-, rhodamine-, aminoallyl-,fluorescein-labeled nucleotides)标准第一链cDNA的合成

 

使用者需要事先准备:

 · dNTP Mix (10mM)

           · Nuclease-Free Water

           · Primer

           · RNA Templete

 

1.  一个常规的反应需要1ng - 5μg total RNA或1ng - 1μg mRNA,

①RNA变性

按照如下配比将各组分加入到已除酶的微量离心管中:

Oligo(dT)12-18 (10pmol) or 25pmol random

primers or 10 pmole gene-specific primer (GSP)      1 μl

dNTP (10mM)                                  2 μl

Nuclease-Free Water                           11-x μl

RNA (1μg/μl)                                   x μl

65度5min,降温到4度

 

②上述变性后退火反应液14ul中加入以下:

M-MLV Revertase 5X Reaction Buffer               4 μl

RNase inhibitor (Rnasin)                          1 μl

M-MLV Revertase                               1 μl

Total                                     20 μl

2.  混匀各组分并短暂温和离心;

 

3.  (此步使用随机引物时必选)30℃ 温浴10min;

 

4.  接着42℃ 温浴30-60min;

 

5.  95℃ 温浴5min终止反应;(长片段77度15min)

 

6.  4℃ 放置5min。

建议:每50 μl PCR反应容积中,加入2-4 μl 第一链cDNA作为模板。

 

注意

1.       PCR产物>1kb的扩增反应要求去除与cDNA互补的RNA,加入1 μl

 (2 units) RNase H37℃ 温浴20min。

2.       逆转录时添加1M betaine (终浓度) 5% DMSO (终浓度)可以增强高GC含量RNA模板的效率。

 

 

应用

· 第一链cDNA合成

· RT-PCR和实时RT-PCR

· 用于克隆和表达的cDNA合成

· 被标记的cDNA探针检测基因芯片

· DNA标记

· RNA分析中的引物延伸

活力单位定义

一个单位活力定义为,42,以poly(A)-oligo(dT)25作为模板-引物,10min内催化1 nmol脱氧核苷酸[3H]dTTP渗入到酸沉淀物中所需酶量。

质量控制

l  经SDS-PAGE电泳分析,纯度达到95%以上;

l  无DNA内切酶、3’和5’端DNA外切酶及RNA酶的活性。

l  标准反应体系中由RNA ladder(分子量范围从1.2kb到7.5kb)合成cDNA,能够得到单一的全长1.2kb的产物和部分全长7.5kb的产物。

 

   

M10511 (10000U)

M10512 (50000U)

M-MLV Revertase

50μl         (200U/μl)

250μl      (200U/μl)

M-MLV Revertase 5× Reaction Buffer

200μl

1ml

   

360

1440

 

 

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