TNF-alpha Recombinant monoclonal antibody, PBS Only (Detector)

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  Sandwich ELISA sample test of MP00702-4

  Human peripheral blood mononuclear cells (PBMC) were cultured unstimulated or stimulated with 10 μg/mL PHA for 3 days. The mean TNF-alpha concentration was determined to be 1.3 pg/mL in unstimulated PBMC supernatant,1236.3 pg/mL in PHA stimulated PBMC supernatant.

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  Sandwich ELISA sample test of MP00702-4

  Serum of seven individual healthy human donors was measured. The human TNF-alpha concentration of detected samples was determined to be 3.25 pg/mL with a range of ND-9.1 pg/mL.

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  Sandwich ELISA standard curve of MP00702-4

  Sandwich ELISA standard curve of MP00702-4, TNF-alpha Recombinant Matched Antibody Pair - PBS only. 83721-5-PBS was coated to a plate as the capture antibody and incubated with serial dilutions of standard Eg0816. 83721-6-PBS was HRP conjugated as the detection antibody. Range: 3.9-250 pg/mL.

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  FC experiment of human PBMCs using 83721-6-PBS

  1x10^6 untreated or PMA, Ionomycin and Brefeldin A treated human PBMCs were intracellularly stained with 0.25 ug TNF-alpha Recombinant monoclonal antibody, PBS Only (Detector) (83721-6-PBS, Clone: 240668A4) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2). Cells were co-stained with 0.25 ug CoraLite® Plus 647 Anti-Human CD4 (RPA-T4) (CL647-65143, Clone: RPA-T4). Cells were fixed and permeabilized with Intracellular Flow Cytometry Fixation & Permeabilization Buffer Kit (PF00019).

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  Affinity and Kinetic Characterization of 83721-6-PBS

  Biolayer interferometry (BLl) kinetic assay of 83721-6-PBS against Human TNF-alpha was performed. The affinity constant is 47.9 pM.

TNF-alpha Uni-rAb^® Recombinant Antibody for FC (Intra), Sandwich ELISA, Indirect ELISA

Cat No. 83721-6-PBS

Clone No.240668A4

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

FC (Intra), Sandwich ELISA, Indirect ELISA

Synonyms

TNF, TNF alpha, TNF-a, C-domain 1, C-domain 2

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Tested Applications

• Positive FC (Intra) detected in: PMA, Ionomycin and Brefeldin A treated human PBMCs

Recommended dilution

• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Product Information

83721-6-PBS targets TNF-alpha as part of a matched antibody pair:

MP00702-4: 83721-5-PBS capture and 83721-6-PBS detection (validated in Sandwich ELISA)

Unconjugated rabbit recombinant monoclonal antibody in PBS only (BSA and azide free) storage buffer at a concentration of 1 mg/mL, ready for conjugation. Created using Proteintech’s proprietary in-house recombinant technology. Recombinant production enables unrivalled batch-to-batch consistency, easy scale-up, and future security of supply.

This conjugation ready format makes antibodies ideal for use in many applications including: ELISAs, multiplex assays requiring matched pairs, mass cytometry, and multiplex imaging applications.Antibody use should be optimized by the end user for each application and assay.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen:

  CatNo: Eg0816

  Product name: Recombinant Human TNF-alpha protein (His Tag)

  Source: mammalian cells-derived, pHZ-KIsec-C-6*HIS

  Tag: C-6*HIS

  Domain: 77-233 aa of NM_000594.4

  Sequence: VRSSSRTPSDKPVAHVVANPQAEGQLQWLNRRANALLANGVELRDNQLVVPSEGLYLIYSQVLFKGQGCPSTHVLLTHTISRIAVSYQTKVNLLSAIKSPCQRETPEGAEAKPWYEPIYLGGVFQLEKGDRLSAEINRPDYLDFAESGQVYFGIIAL

• Full Name: tumor necrosis factor (TNF superfamily, member 2)
• Calculated Molecular Weight: 26 kDa
• GenBank Accession Number: NM_000594.4
• Gene Symbol: TNF-alpha
• Gene ID (NCBI): 7124
• ENSEMBL Gene ID: ENSG00000232810
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• UNIPROT ID: P01375
• Storage Buffer: PBS only, pH 7.3.
• Storage Conditions: Store at -80°C.

Background Information

TNF, as also known as TNF-alpha, or cachectin, is a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. It is expressed as a 26 kDa membrane bound protein and is then cleaved by TNF-alpha converting enzyme (TACE) to release the soluble 17 kDa monomer, which forms homotrimers in circulation. It is produced chiefly by activated macrophages, although it can be produced by many other cell types such as CD4+ lymphocytes, NK cells, neutrophils, mast cells, eosinophils, and neurons. It can bind to, and thus functions through its receptors TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. This cytokine is involved in the regulation of a wide spectrum of biological processes including cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, ins resistance, and cancer.