Product Pathways - Growth Factors/Cytokines
TNF-α (D1G2) Rabbit mAb (IF/Flow Preferred) #8184
PhosphoSitePlus® protein, site, and accession data: TNF-a
| Applications | Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|
| W IP IF-IC F | H | Endogenous | 18, 25 | Rabbit |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
Specificity / Sensitivity
TNF-α (D1G2) Rabbit mAb (IF/Flow Preferred) recognizes endogenous levels of total TNF-α protein. TNF-α (D5G9) Rabbit mAb #6945 is more sensitive by western blot.
Source / Purification
Monoclonal antibody is produced by immunizing animals with recombinant human TNF-α protein.
Western Blotting
Western blot analysis of extracts from THP-1 cells, differentiated with TPA #4174 (80 nM, overnight) and then untreated (-) or treated with LPS (1 μg/ml, 1 hr; +), using TNF-α (D1G2) Rabbit mAb (IF/Flow Preferred) (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Flow Cytometry
Flow cytometric analysis of human peripheral blood lymphocytes, untreated (left) or treated with TPA #4174 (40 nM, 4 hr), Ionomycin #9995 (2 μM, 4 hr), and Brefeldin A #9972 (1 μg/ml, last 3 hr of treatment) (right), using a CD3 antibody and TNF-α (D1G2) Rabbit mAb (IF/Flow Preferred).
IF-IC
Confocal immunofluorescent analysis of THP-1 cells, treated with TPA #4174 (80 nM, 16 hr) (left) or treated with TPA followed by LPS (1 μg/ml, 1 hr) (right), using TNF-α (D1G2) Rabbit mAb (IF/Flow Preferred) (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
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, Erk1/2, 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).
- Aggarwal, B.B. (2003) Nat Rev Immunol 3, 745-56.
- Hehlgans, T. and Pfeffer, K. (2005) Immunology 115, 1-20.
- Lin, P.L. et al. (2007) J Investig Dermatol Symp Proc 12, 22-5.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 7321 Human TNF-α Neutralizing (D1B4) Rabbit mAb
- 6945 TNF-α (D5G9) Rabbit mAb
- 3707 TNF-α Antibody
- 8902 Human Tumor Necrosis Factor-α (hTNF-α)
For Research Use Only. Not For Use In Diagnostic Procedures.
