Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Human Latent Transforming Growth Factor β1 (hLatent TGF-β1) #5154

Latency-associated peptide   TGF-B1 + LAP  

No. Size Price
5154LC 10 µg ( With Carrier ) ¥2,580.00 现货查询 购买询价
5154LF 10 µg ( Carrier Free ) ¥2,580.00 现货查询 购买询价
5154 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype

Species cross-reactivity is determined by western blot.

Applications Key:

Source / Purification

Recombinant human latent TGF-β1 (hLatent TGF-β1) Leu30-Ser390 (Accession #P01137) was expressed in human 293 cells at Cell Signaling Technology.

Cell Signaling Technology使用人293T细胞表达重组人latent TGF-β1 (hLatent TGF-β1) Leu30-Ser390 (Accession #P01137)。

Molecular Characterization

Recombinant hLatent TGF-β1 contains no "tags" and the nonglycosylated small latent TGF-β1 complex has a calculated MW of 41,251. DTT-reduced protein migrates as 40 and 13 kDa polypeptides, and the non-reduced cystine-linked homodimers migrate as 80 and 25 kDa proteins. The expected amino-terminal ALDTN of recombinant hTGF-β1 and the expected amino-terminal LSTSK of recombinant latency-associated peptide (LAP) were verified by amino acid sequencing.

重组hLatent TGF-β1没有标签,计算得到非糖基化的小潜在TGF-β1复合物分子量为41,251。DTT-还原蛋白迁移率显示其包含两条多肽分别为40和13kDa,非还原状态以半胱氨酸连接形成的同二聚体显示蛋白分别为80和25kDa。重组hTGF-β1氨基端序列ALDTN和重组LAP氨基端序列LSTSK通过氨基酸测序确认。

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hLatent TGF-β1. All lots are greater than 98% pure.还原和未还原的mEGF由SDS-PAGE测定纯度大于98%。所有批次的产品纯度均大于98%。

Bioactivity

The bioactivity of recombinant hLatent TGF-β1 was determined by assessing inhibition of IL-4 induced HT-2 cell proliferation. The ED50 of each lot is between 4-10 ng/ml and 0.2-0.8 ng/ml after acid activation.

通过抑制IL-4诱导HT-2细胞增殖的实验,以检测重组蛋白hLatent TGF-β1的生物活性。各批次产品经酸激活后ED50在4-10 ng/ml 和 0.2-0.8 ng/ml之间。

Coomassie Gel

Coomassie Gel

The purity of recombinant hLatent TGF-β1 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hLatent TGF-β1 and staining overnight with Coomassie Blue.重组人hLatent TGF-β1 的纯度使用SDS-PAGE确认,6 ug还原(+)和未还原(-)的重组hLatent TGF-β1 电泳后使用考马斯蓝染色过夜。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or treated with the hLatent TGF-β1 for 25 minutes, using Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb #3108 (upper) and Smad2 (86F7) Rabbit mAb #3122 (lower).使用人hLatent TGF-β1处理25分钟或不处理的HeLa细胞提取物进行Western blot实验,用Phospho-Smad2 (Ser465/467) (138D4) 兔源单克隆抗体#3108 (上)和Smad2(86F7)兔源单克隆抗体#3122(下)进行检测。

Bioactivity

Bioactivity

The inhibition of IL-4 induced proliferation in HT-2 cells treated with increasing concentrations of hLatent TGF-β1 or acid-activated hLatent TGF-β1 was assessed. After 48 hour treatment with hLatent TGF-β1, cells were incubated with a chemiluminescent cell viability reagent and the relative light units (RLU) were determined.逐渐增长浓度的 hLatent TGF-β1 或酸激活的 hLatent TGF-β1 抑制IL-4 诱导的HT-2细胞的增殖的能力通过实验验证。hLatent TGF-β1处理48小时候,使用一种化学荧光活性试剂抚育细胞,随后检测相对光量(RLU)。

Endotoxin

Less than 0.01 ng endotoxin/1 μg hLatent TGF-β1.每1ug hLatent TGF-β1所含内毒素低于0.01ng。

Formulation

With carrier: A 0.22 μm filtered solution of 0.25 mg/ml hLatent TGF-β1 in PBS, pH 7.2 and 25% (v/v) glycerol containing 20 μg BSA per 1 μg hLatent TGF-β1. Carrier free: A 0.22 μm filtered solution of 0.25 mg/ml hLatent TGF-β1 in PBS, pH 7.2 and 25% (v/v) glycerol.带载体: 0.25 mg/ml hLatent TGF-β1溶于pH 7.2,25% (v/v) glycerol 的PBS溶液使1 μg hLatent TGF-β1携带20 μg BSA,溶液经0.22um滤器过滤获得。无载体:0.25 mg/ml hLatent TGF-β1溶于含有25% (v/v) glycerol pH 7.2 的PBS溶液经0.22um滤器过滤获得。

Background

Latent TGF-β1 is a complex of two proteins, latency associated protein (LAP) and TGF-β1, which is derived from cleavage of a common 75 kDa precursor protein (1). The LAP protein spatially and temporally regulates TGF-β1 activity by sequestering TGF-β1 in the extracellular matrix in conjunction with latent TGF-β1 binding proteins (LTBP)(1). The release of TGF-β1 is activated by a number of stimuli including proteases, thrombospondin-1, reactive oxygen species, and some integrins (1). Active TGF-β1 binds to TβRII homodimer, which then complexes with TβRI homodimer (2,3). The oligomeric receptor complex phosphorylates subsets of the Smad proteins that then act to induce or repress a number of target genes (3-5). TGF-β1 binding can also activate the Erk2, p38, and Jnk pathways via TAK1 (5). Active TGF-β1 activities include proliferation, angiogenesis, and promotion or inhibition of many immune events (2,4,5). Latent TGF-β1 is present on the surface of regulatory T cells in association with GARP and may contribute directly to their immunosuppressive activity (6,7).

潜伏的TGF-β1是两个蛋白组成的复合物,潜伏相关蛋白(LAP)和TGF-β1,TGF-β1由一个75 kDa的前体蛋白的剪切后得到(1)。LAP蛋白可以时空特异性的调控TGF-β1,它促进TGF-β1蛋白与潜在TGF-β1结合蛋白(LTBP)结合,使其留在细胞外基质中(1)。一系列的刺激包括蛋白酶,凝血酶敏感蛋白1,活性氧和一些整合素都可以激活TGF-β1的释放(1)。有活性的TGF-β1与TβRII同二聚体结合,随后再与TβRI同二聚体形成复合物(2,3)。这种低聚受体复合物可以磷酸化Smad蛋白以激活或抑制多种靶基因表达(3-5)。TGF-β1的结合也可以通过TAK1激活Erk2, p38, 和Jnk信号通路(5)。有活性的TGF-β1的功能包括增殖,血管生成和促进或抑制许多免疫反应(2,4,5)。潜伏的TGF-β1与GARP关联由调节T细胞呈递至细胞表面,可能与免疫抑制反应直接相关(6,7)。

Application References

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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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