Cell-based Screening Assays - Ozyme

Cytokines and Growth Factors
The world’s highest quality antibody provider has now extended its expertise to cytokine and growth factor production.
Cell Signaling Technology (CST) is now offering a
selection of cytokines and growth factors. These
reagents are produced and bioassayed in-house, and
are held to the same unparalleled quality standards as
the CST antibodies you know and trust.
:: Produced and bioassayed in-house with the
highest purity and bioactivity.
:: Comparison of multiple lots, stringent product
specifications, and rigorous quality control
ensures maximum lot-to-lot consistency.
:: Endotoxin levels are routinely tested and are
less than 0.01 ng/µg cytokine.
:: Many products are produced in mammalian
cells to maximize natural conformation and
glycosylation.
:: Multi-milligram quantities available.
Visit our website for the most up-to-date
product listing.
Lot-to-Lot Consistency
A
B
OD450-OD650
0.6
0.3
0
kDa
200
116
97
66
55
37
31
22
14
6
4
Lot 1 Lot 2 Lot 3
0.6
0.6
0.01 0.1 1 10 100
hIL-6 (ng/ml)
+
hIL-6
OD450-OD650
0.3
0
kDa
200
116
97
66
55
37
31
22
14
6
4
0.01 0.1 1 10 100
hIL-6 (ng/ml)
+
0.01 0.1 1 10 100
hIL-6 (ng/ml)
Comparison of purity and bioactivity of three independent lots of Human Interleukin-6 (hIL-6) #8904:
The proliferation of TF-1 cells was assessed after 48 hr treatment with increasing concentrations of hIL-6. Cells were incubated with a
tetrazolium salt and the OD 450 - OD 650 was determined (A). The purity of recombinant hIL-6 was determined by SDS-PAGE using 6 µg
reduced (+) and non-reduced (-) recombinant hIL-6 and staining overnight with Coomassie Blue (B).
hIL-6
OD450-OD650
0.3
0
kDa
200
116
97
66
55
37
31
22
14
6
4
+
hIL-6
Human Tumor Necrosis Factor-α (hTNF-α) #8902 validation includes:
A Purity
B Bioactivity
C Downstream Signaling,
Western Blot
D
kDa
116
97
66
55
37
31
22
14
6
3
+ –
hTNF-α
OD 595 nm
0.4
0.2
0
0 .01 .1 1 10 100
hTNF-α (ng/ml)
kDa
100
80
60
50
100
80
60
50
–
0.01
0.1
1
10
Phospho-NF-κB
p65 (Ser536)
(93H1) Rabbit mAb
#3033
NF-κB p65
Antibody
#3034
hTNF-α (ng/ml)
Human Tumor Necrosis Factor-α (hTNF-α) #8902: (A) 6 µg reduced (+) and non-reduced (-) hTNF-α; hTNF-α does not contain tags or additional
amino acids. (B) The viability of L-929 cells treated with increasing amounts of hTNF-α in the presence of 2 ng/ml actinomycin D was determined. (C)
Phosphorylation of NF-κB p65 was measured in response to treatment with hTNF-α. (D) HT-1080 cells were treated with increasing concentrations
of hTNF-α to assess the effect on nuclear translocation of NF-κB, detected using NF-κB p65 (D14E12) XP ® Rabbit mAb #8242. All data points were
performed in triplicate to ensure validity and reproducibility of the assay. The signal was analyzed using an Cellomics ArrayScan ® V TI . Inset: Confocal
IF analysis of HT-1080 cells, untreated (left) or treated with hTNF-α (20 ng/ml, 20 min) (right), using NF-κB p65 (D14E12) XP ® Rabbit mAb #8242
(green). Red = Actin filaments (DY-554 phalloidin); Blue = DRAQ5 ® #4084 (DNA dye).
Normalized
Nuclear Fluorescence Intensity
170
160
150
140
130
120
110
100
Downstream Signaling,
Immunofluorescent Analysis
untreated
hTNF-α-treated
0.001 0.01 0.1
hTNF-α (ng/ml)
1 10
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