Recombinant human TNF alpha protein (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
Description
-
Product name
Recombinant human TNF alpha protein
See all TNF alpha proteins and peptides -
Biological activity
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.
-
Purity
> 98 % SDS-PAGE.
>98% by HPLC analyses. Sterile filtered. -
Endotoxin level
< 1.000 Eu/µg -
Expression system
Escherichia coli -
Accession
-
Protein length
Full length protein -
Animal free
No -
Nature
Recombinant -
-
Species
Human -
Sequence
VRSSSRTPSD KPVAHVVANP QAEGQLQWLN RRANALLANG VELRDNQLVV PSEGLYLIYS QVLFKGQGCP STHVLLTHTI SRIAVSYQTK VNLLSAIKSP CQRETPEGAE AKPWYEPIYL GGVFQLEKGD RLSAEINRPD YLDFAESGQV YFGIIAL -
Predicted molecular weight
17 kDa -
Amino acids
77 to 233 -
Additional sequence information
aa 77 to 233 refers to the full length mature form (soluble).
-
Associated products
-
ELISA pair antibody
-
Related Products
Specifications
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.
-
Applications
Functional Studies
Sandwich ELISA
HPLC
SDS-PAGE
-
Form
Lyophilized -
Additional notes
Lots prior to June 2015 contain 0.036% Tris (including lots GR157466-11, -12 and -13).
-
Concentration information loading...
Preparation and Storage
-
Stability and Storage
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.
-
ReconstitutionReconstitute with dH2O to make a final concentration between 0.1 to 1.0 mg/ml.
General Info
-
Alternative names
- APC1
- APC1 protein
- Cachectin
see all -
Function
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. -
Involvement in disease
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). -
Sequence similarities
Belongs to the tumor necrosis factor family. -
Post-translational
modificationsThe 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. -
Cellular localization
Secreted and Cell membrane. - Information by UniProt
Images
-
Sandwich ELISA - Recombinant human TNF alpha protein (ab9642)
-
ChIP - Recombinant human TNF alpha protein (ab9642)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).
-
Western blot - Recombinant human TNF alpha protein (ab9642)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? -
Western blot - Recombinant human TNF alpha protein (ab9642)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).
-
Western blot - Recombinant human TNF alpha protein (ab9642)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.
-
Immunoprecipitation - Recombinant human TNF alpha protein (ab9642)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.
-
Western blot - Recombinant human TNF alpha protein (ab9642)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.
Protocols
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.
Datasheets and documents
-
Datasheet download
References (20)
ab9642 has been referenced in 20 publications.
- 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
