Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Human Tumor Necrosis Factor-α (hTNF-α) #8902

cachectin   ikb   IκB-α   IκBα   lymphotoxin   nf-kappab   nf-kb   nfkb   rel   tnf   tnf-a   tnf-alpha   tnfa   tnfr   traf   tumor necrosis factor alpha   tumour necrosis factor alpha  

No. Size Price
8902LC 50 µg ( With Carrier ) ¥6,860.00 现货查询 购买询价
8902LF 50 µg ( Carrier Free ) ¥6,860.00 现货查询 购买询价
8902SC 10 µg ( With Carrier ) ¥2,321.00 现货查询 购买询价
8902SF 10 µg ( Carrier Free ) ¥2,321.00 现货查询 购买询价
8902 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 TNF-α (hTNF-α) Val77-Leu233 (Accession #HUMTNFAB) was produced in E. coli at Cell Signaling Technology.

在CST公司,重组的人TNF-α (hTNF-α) Val77-Leu233 (Accession #HUMTNFAB)是在E. coli中表达产生。

Molecular Characterization

Recombinant hTNF-α does not have a Met on the amino terminus and has a calculated MW of 17,352. DTT-reduced and non-reduced protein migrate as 18 kDa polypeptides. The expected amino-terminal VRSSS of recombinant hTNF-α was verified by amino acid sequencing. TNF-α is a non-disulfide-linked homotrimer in solution as determined by chemical cross-linking.

重组的hTNF-α蛋白在N-末端无 Met标签,其分子量据推算为17,352Da。DTT-还原和未还原的蛋白迁移时是作为一个18kDa 的多肽而转移。重组hTNF-α蛋白的N-末端序列VRSSS通过测序得到。hTNF-α在溶液中是无二硫键连接的三聚物,而依靠化学连接。

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hTNF-α. All lots are greater than 98% pure.

6 μg 还原 (+) 和 非还原的(-) hTNF-α通过SDS-PAGE检测,纯度大于98%,所有批次的纯度均高于98%。

Bioactivity

The bioactivity of hTNF-α was determined in an L-929 cell viability assay. The ED50 of each lot is between 10-100 pg/ml.

hTNF-α的生物活性是通过L-929细胞生存能力实验确定的。每个批次的ED50在10-100 pg/ml之间。

Coomassie Gel

Coomassie Gel

The purity of recombinant hTNF-α was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hTNF-α and staining overnight with Coomassie Blue.重组蛋白hTNF-α的纯度用6 µg 还原 (+) 和未还原(-)的蛋白走SDS-PAGE 胶来观察。考马斯亮蓝染色过夜。

Bioactivity

Bioactivity

The viability of L-929 cells treated with increasing amounts of hTNF-α in the presence of 2 ng/ml actinomycin D was determined. Cells were stained with crystal violet at the end of treatment and the OD595 was determined.L-929细胞的生存能力,在2 ng/ml放射菌素D 存在下用递增量的hTNF-α处理。处理结束后细胞用结晶紫染色并测定OD595数值。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells treated with hTNF-α for 20 minutes, using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb #3033 (upper) and total NF-κB p65 Antibody #3034 (lower).Western免疫印迹。用Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb #3033(上图)和total NF-κB p65 Antibody #3034(下图)研究经hTNF-α处理20分钟的HeLa细胞的细胞提取液。

Endotoxin

Less than 0.01 ng endotoxin/1 μg hTNF-α.

内毒素含量:<0.01 ng /1 μghTNF-α。

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg hTNF-α. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.

有载体: 每微克hTNF-α蛋白溶解在包含 20 μg BSA的PBS(pH 7.2)溶液中,并通过 0.22 μm 滤膜冻干。

无载体:每微克hTNF-α蛋白溶解在PBS(pH 7.2)溶液中,并通过 0.22 μm 滤膜冻干。

Background

TNF-α, the prototypical member of the TNF protein superfamily, is a homotrimeric type-II membrane protein (1,2). Membrane bound TNF-α is cleaved by the metalloprotease TACE/ADAM17 to generate a soluble homotrimer (2). Both membrane and soluble forms of TNF-α are biologically active. TNF-α is produced by a variety of immune cells including T cells, B cells, NK cells and macrophages (1). Cellular response to TNF-α is mediated through interaction with receptors TNF-R1 and TNF-R2 and results in activation of pathways that favor both cell survival and apoptosis depending on the cell type and biological context. Activation of kinase pathways (including JNK, ERK (p44/42), p38 MAPK and NF-κB) promotes the survival of cells, while TNF-α mediated activation of caspase-8 leads to programmed cell death (1,2). TNF-α plays a key regulatory role in inflammation and host defense against bacterial infection, notably Mycobacterium tuberculosis (3). The role of TNF-α in autoimmunity is underscored by blocking TNF-α action to treat rheumatoid arthritis and Crohn’s disease (1,2,4).

TNF-α, TNF家族的典型成员, 是II型的同源三聚体膜蛋白(1,2)。细胞膜锚定的TNF-α通过金属蛋白酶TACE/ADAM17的剪切形成可溶性的同型三聚体(2)。膜锚定和可溶性两种形式都有生物活性。TNF-α可由很多种类的细胞产生包括T细胞、B细胞、巨噬细胞和NK细胞(1)。TNF-α的细胞内响应是通过与 TNF-R1 和 TNF-R2 两个受体互作而介导的并激活细胞存活和细胞凋亡两个截然相反地过程,这两个过程主要决定于细胞的类型和生物学环境。激活激酶的通路(包括JNK, Erk (p44/42), p38 MAPK 和 NF-κB) 促使细胞的存活,TNF-α介导的对caspase-8的激活导致了程序性的细胞死亡(1,2)。TNF-α在炎症反应和宿主抵抗细菌入侵的过程特别是结核分枝杆菌发挥了重要的调控作用(3)。在通过抑制TNF-α来治疗类风湿性关节炎和Crohn’s disease的过程中,TNF-α在自免疫中的作用被低估(1,2,4)。

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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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