Store at -20°C
TNF-α Antibody
#3707
3 100 µl
n
Orders n 877-616-CELL (2355)
orders@cellsignal.com
Support n 877-678-TECH (8324)
info@cellsignal.com
Web n www.cellsignal.com
(10 western blots)
rev. 05/20/15
For Research Use Only. Not For Use In Diagnostic Procedures.
Entrez-Gene ID #7124
Swiss-Prot Acc. #P01375
Applications
Species Cross-Reactivity*
Molecular Wt.
Source
W, IP
Endogenous
H, M, (Mk, Pg)
17 kDa mature
25 kDa precursor
Rabbit**
Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM
NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C.
Do not aliquot the antibody.
*Species cross-reactivity is determined by western blot.
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).
Recommended Antibody Dilutions:
Western Blottin
1:1000
Immunoprecipitation1:25
200
140
100
80
For product specific protocols and a complete listing
of recommended companion products please see the
product web page at www.cellsignal.com
60
50
40
30
20
TNF-α
10
1
5
10
TNF-α (ng)
Western blot analysis of recombinant Human Tumor Necrosis
Factor-α (hTNF-α) #8902 using TNF-α Antibody.
Specificity/Sensitivity: TNF-a Antibody detects endogenous levels of mature TNF-a as well TNF-a precursor
protein.
© 2015 Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
**Anti-rabbit secondary antibodies must be used to
detect this antibody.
kDa
kDa
200
140
100
80
Source/Purification: Polyclonal antibodies are produced
by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly198 of human TNF-α protein.
Antibodies are purified by protein A and peptide affinity
chromatography.
60
50
40
Background References:
(1) Aggarwal, B.B. (2003) Nat Rev Immunol 3, 745-56.
30
(2) Hehlgans, T. and Pfeffer, K. (2005) Immunology 115,
1-20.
20
TNF-α
(3) Lin, P.L. et al. (2007) J Investig Dermatol Symp Proc
12, 22-5.
–
+ LPS
Western blot analysis of extracts from Raw 264.7 cells, untreated
(-) or LPS-treated (1 µg/ml, 18 hr; +), using TNF-α Antibody.
(4) Brennan, F.M. and McInnes, I.B. (2008) J Clin Invest
118, 3537-45.
IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS,
0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key:
W—Western
Species Cross-Reactivity Key:
IP—Immunoprecipitation
H—human
M—mouse
Dg—dog Pg—pig Sc—S. cerevisiae Ce—C. elegans
IHC—Immunohistochemistry
R—rat
Hr—Horse
Hm—hamster
ChIP—Chromatin Immunoprecipitation
Mk—monkey
All—all species expected
Mi—mink
C—chicken
Tween is a registered trademark of ICI Americas, Inc.
IF—Immunofluorescence
F—Flow cytometry
Dm—D. melanogaster X—Xenopus
Z—zebrafish
Species enclosed in parentheses are predicted to react based on 100% homology.
E-P—ELISA-Peptide
B—bovine