重组人TNF alpha蛋白(ab9642)
Key features and details
- Expression system: Escherichia coli
- Purity: > 98% SDS-PAGE
- Endotoxin level: < 1.000 Eu/µg
- Active: Yes
- Suitable for: Functional Studies, Sandwich ELISA, HPLC, SDS-PAGE
Achieve higher consistency and quality standards with a premium grade bioactive protein
- High batch-to-batch consistency
- Optimal bioactivity
- Guaranteed identical to human native proteins
- >95% purity
- Ultra-low endotoxin levels: <0.005 Eu/µg
- Carrier and tag free
描述
-
产品名称
重组人TNF alpha蛋白
参阅全部 TNF alpha 蛋白酶 -
生物活性
The ED50, as determined by the cytolysis of murine L929 cells in the presence of Actinomyocin D, is ≤ 0.05 ng/mL, corresponding to a specific activity of ≥ 2 x 107 units/mg.
-
纯度
> 98 % SDS-PAGE.
>98% by HPLC analyses. Sterile filtered. -
内毒素水平
< 1.000 Eu/µg -
表达系统
Escherichia coli -
Accession
-
蛋白长度
Full length protein -
无动物成分
No -
性质
Recombinant -
-
种属
Human -
序列
VRSSSRTPSD KPVAHVVANP QAEGQLQWLN RRANALLANG VELRDNQLVV PSEGLYLIYS QVLFKGQGCP STHVLLTHTI SRIAVSYQTK VNLLSAIKSP CQRETPEGAE AKPWYEPIYL GGVFQLEKGD RLSAEINRPD YLDFAESGQV YFGIIAL -
预测分子量
17 kDa -
氨基酸
77 to 233 -
额外的序列信息
aa 77 to 233 refers to the full length mature form (soluble).
-
相关产品
-
ELISA pair antibody
-
Related Products
技术指标
Our Abpromise guarantee covers the use of ab9642 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
-
应用
Functional Studies
Sandwich ELISA
HPLC
SDS-PAGE
-
形式
Lyophilized -
补充说明
Lots prior to June 2015 contain 0.036% Tris (including lots GR157466-11, -12 and -13).
-
Concentration information loading...
制备和贮存
-
稳定性和存储
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Constituents: 0.049% Sodium phosphate, 0.12% Sodium chloride
This product is an active protein and may elicit a biological response in vivo, handle with caution.
-
复溶Reconstitute with dH2O to make a final concentration between 0.1 to 1.0 mg/ml.
常规信息
-
别名
- APC1
- APC1 protein
- Cachectin
see all -
功能
Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation. -
疾病相关
Genetic variations in TNF are a cause of susceptibility psoriatic arthritis (PSORAS) [MIM:607507]. PSORAS is an inflammatory, seronegative arthritis associated with psoriasis. It is a heterogeneous disorder ranging from a mild, non-destructive disease to a severe, progressive, erosive arthropathy. Five types of psoriatic arthritis have been defined: asymmetrical oligoarthritis characterized by primary involvement of the small joints of the fingers or toes; asymmetrical arthritis which involves the joints of the extremities; symmetrical polyarthritis characterized by a rheumatoidlike pattern that can involve hands, wrists, ankles, and feet; arthritis mutilans, which is a rare but deforming and destructive condition; arthritis of the sacroiliac joints and spine (psoriatic spondylitis). -
序列相似性
Belongs to the tumor necrosis factor family. -
翻译后修饰
The soluble form derives from the membrane form by proteolytic processing.
The membrane form, but not the soluble form, is phosphorylated on serine residues. Dephosphorylation of the membrane form occurs by binding to soluble TNFRSF1A/TNFR1.
O-glycosylated; glycans contain galactose, N-acetylgalactosamine and N-acetylneuraminic acid. -
细胞定位
Secreted and Cell membrane. - Information by UniProt
图片
-
Chromatin was prepared from HeLa (human epithelial cell line from cervix adenocarcinoma) cells treated with and without 20 ng/ml TNF-α (ab9642) for 60 minutes according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25 μg of chromatin, 5 μg of ab218533 (red), and 20 µl of Protein A/G sepharose beads. 5 μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (SYBR green approach).
The ChIP data are consistent with the literature (PMID: 16135789).
-
All lanes : Anti-TNFAIP3 antibody [EPR2663] (ab92324) at 1/5000 dilution
Lane 1 : WEHI-3 (Mouse leukemia lymphoblast) whole cell lysate
Lane 2 : WEHI-3 treated with 20 ng/ml TNF alpha (ab9642) for 6 h
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 80 kDa why is the actual band size different from the predicted? -
All lanes : Anti-NF-kB p65 (acetyl K310) antibody [EPR21781] - ChIP Grade (ab218533) at 1/2000 dilution
Lane 1 : HEK-293 transfected with NF-kB p65 expression vector containing a myc-His-tag®, whole cell lysate
Lane 2 : HEK-293 transfected with NF-kB p65 and p300 (aa1287-1663) expression vectors containing a myc-His-tag®, then treated with 20 ng/ml TNF-alpha (ab9642) for 60 minutes, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Observed band size: 70 kDa why is the actual band size different from the predicted?
Exposure time: 37 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
NF-κB p65 (acetyl K310) expression is induced by TNF-α and p300 acetyltransferases (PMID: 20160011, PMID: 12456660, PMID: 16135789).
-
All lanes : Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436) at 1/1000 dilution
Lane 1 : Untreated L-929 (Mouse connective tissue fibroblast cells) whole cell lysate
Lane 2 : L-929 whole cell lysate treated with 20 ng/ml TNF alpha (ab9642), 100 nM Smac mimetic, and 20 µM z-VAD (ab120382) for 8 h and then harvested.
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Observed band size: 54 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking and dilution buffer: 5% NFDM/TBST.
-
MLKL (phospho S345) was immunoprecipitated from 1mg of L-929 (Mouse connective tissue fibroblast cells) whole cell lysate treated with 20 ng/ml TNF alpha (ab9642) + 100 nM Smac mimetic + 20 µM z-VAD compound (ab120382) for 8h using ab196436 at 1/150 dilution. Western blot was performed from the immunoprecipitate using ab196436 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: L-929 whole cell lysate treated with 20 ng/ml TNF alpha (ab9642) + 100 nM Smac mimetic+ 20 µM z-VAD compound (ab120382) for 8h;10 µg (Input).
Lane 2: ab196436 IP in L-929 whole cell lysate treated with 20 ng/ml TNF alpha (ab9642) + 100 nM Smac mimetic+ 20 µM z-VAD compound (ab120382) for 8h.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab196436 in L-929 whole cell lysate treated with 20 ng/ml TNF alpha (ab9642) + 100 nM Smac mimetic+ 20 µM z-VAD compound (ab120382) for 8h.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
-
All lanes : Anti-MLKL (phospho S345) antibody [EPR9515(2)] (ab196436) at 1/1000 dilution
Lane 1 : L-929 treated with 20 ng/ml TNF alpha (ab9642), 100 nM Smac mimetic, and 20 µM z-VAD (ab120382) for 8 h, whole cell lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Mouse colon tissue lysate
Lane 4 : Mouse lung tissue lysate
Lane 5 : Mouse retina tissue lysate
Lane 6 : Mouse liver tissue lysate
Lane 7 : Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 54 kDa why is the actual band size different from the predicted?
Exposure time: 50 secondsBlocking and diluting buffer: 5% NFDM/TBST.
MLKL pS345 is a trigger for necroptosis. It is only detectable in infection/cellular damaged (PMID:29229989) or aging tissue (PMID: 28807105) but not in normal tissues.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
-
Datasheet download
文献 (20)
ab9642 被引用在 20 文献中.
- Zhu L et al. Toxoplasma gondii Rhoptry Protein 7 (ROP7) Interacts with NLRP3 and Promotes Inflammasome Hyperactivation in THP-1-Derived Macrophages. Cells 11:N/A (2022). PubMed: 35626667
- Wang W et al. Engineering micro oxygen factories to slow tumour progression via hyperoxic microenvironments. Nat Commun 13:4495 (2022). PubMed: 35918337
- Wang H et al. Carbon nano-onion-mediated dual targeting of P-selectin and P-glycoprotein to overcome cancer drug resistance. Nat Commun 12:312 (2021). PubMed: 33436622
- Lucas JH et al. Multi-Walled Carbon Nanotubes (MWCNTs) Cause Cellular Senescence in TGF-ß Stimulated Lung Epithelial Cells. Toxics 9:N/A (2021). PubMed: 34205339
- Chitti SV et al. Repurposing of Antibiotic Sulfisoxazole Inhibits Lipolysis in Pre-Clinical Model of Cancer-Associated Cachexia. Biology (Basel) 10:N/A (2021). PubMed: 34439933