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#5358

Sterile

Human Interleukin-10 (hIL-10)

(Mammalian Derived)

New 07/10

n SC 10 µg

(With Carrier)

n LC 50 µg

(With Carrier)

n SF 10 µg

(Carrier Free)

n LF 50 µg

(Carrier Free)

Source: Recombinant human IL-10 (hIL-10) Ser19 -

Asn178 (Accession # NP_000563) was expressed in human

293 cells at Cell Signaling Technology.

Molecular Characterization: Recombinant hIL-10

contains no “tags” and has a calculated MW of 18,647.

DTT-reduced protein migrates as an 18 kDa polypeptide and

non-reduced protein migrates as a 16 kDa polypeptide due

to intramolecular cystines. The expected amino-terminal

SPGQG of recombinant hIL-10 was veriﬁed by amino acid

sequencing.

Endotoxin: Less than 0.01 ng endotoxin/1μg hIL-10.

Purity: >98% as determined by SDS-PAGE of 6 μg

reduced (+) and non-reduced (-) recombinant hIL-10. All

lots are greater than 98% pure.

Bioactivity: The bioactivity of recombinant hIL-10 was

determined in a MC/9 cell proliferation assay. The ED

each lot is between 0.3-1.4 ng/ml.

Formulation: With carrier: Lyophilized from a 0.22 μm ﬁltered

solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg hIL-10.

Carrier free: Lyophilized from a 0.22 μm ﬁltered solution of PBS,

pH 7.2.

Reconstitution:

With carrier: Add sterile PBS, or PBS containing 1% bovine or

human serum albumin or 5-10% FBS to a ﬁnal hIL-10 concen-

tration of greater than 50 μg/ml. Solubilize for 30 minutes at

room temperature with occasional gentle vortexing.

Carrier free: Add sterile PBS, or PBS containing protein to mini-

mize absorption of hIL-10 to surfaces. Solubilize for 30 minutes

at room temperature with occasional gentle vortexing. Stock

hIL-10 should be greater than 50 μg/ml.

Storage: Stable in lyophilized state at 4ºC for 1 year after

receipt. Sterile stock solutions reconstituted with carrier protein

are stable at 4ºC for 2 months and at -20ºC for 6 months. Avoid

repeated freeze-thaw cycles.

Maintain sterility. Storage at -20ºC should be in a manual defrost

freezer.

Applications: Optimal concentration for the desired application

should be determined by the user.

Background: IL-10 is an anti-inﬂammatory cytokine that is pro-

duced by T cells, NK cells and macrophages (1,2). IL-10 initiates

signal transduction by binding to a cell surface receptor complex

consisting of IL-10 RI and IL-10 RII (1). Binding of IL-10 leads

to the activation of Jak1 and Tyk2, which phosphorylates Stat3

(1,3). The anti-inﬂammatory activity of IL-10 is due to its ability

to block signaling through other cytokine receptors, notably

IFNγ receptor, by upregulating expression of SOCS1 (1,3). In

addition, IL-10 promotes T cell tolerance by inhibiting tyrosine

phosphorylation of CD28 (4,5). IL-10 is an important negative

regulator of the immune response, which allows for maintenance

of pregnancy (1). In contrast, increased IL-10 levels contribute to

persistent Leishmania major infections (6).

Background References:

(1) Pestka, S. et al. (2004) Immunol Rev 202, 8-32.

(2) Akuffo, H. et al. (1999) Clin Exp Immunol 117, 529-34.

(3) O’Shea, J.J. and Murray, P.J. (2008) Immunity 28, 477-87.

(4) Akdis, C.A. and Blaser, K. (2001) Immunology 103, 131-6.

(5) Akdis, C.A. et al. (2000) FASEB J 14, 1666-8.

(6) Von Stebut, E. (2000) Eur J Dermatol 17, 115-22.

Orders

877-616-CELL (2355)

[email protected]

Support

877-678-TECH (8324)

[email protected]

Web

www.cellsignal.com

Multi-milligram quantities available

This product is intended for research purposes only. This product is not intended to be used for

therapeutic or diagnostic purposes in humans or animals.

The purity of recombinant hIL-10 was determined by SDS-PAGE

of 6 µg reduced (+) and non-reduced (-) recombinant hIL-10

and staining overnight with Coomassie Blue.

kDa

hIL-10

200

116

+ –

The proliferation of MC/9 cells treated with increasing

concentrations of hIL-10 in the presence of 1 pg/ml mouse IL-4

#5208 was assessed. After 72 hours treatment with hIL-10, cells

were incubated with a tetrazolium salt and the OD

450

-OD

650

was

determined.

0.4

0.3

0.2

0.1

0.001

0.01

0.1

10 100 1000

450

-OD

650

hIL-10 (ng/ml)

Western blot analysis of extracts from RPMI 8226 cells,

untreated or treated with hIL-10 for 10 minutes, using Phospho-

Stat3 (Tyr705) (D3A7) Rabbit mAb #9145 (upper) or Stat3

Antibody #9132 (lower).

kDa

140

100

140

100

200

hIL-10 (ng/ml)

Phospho-Stat3

(Tyr705)

Stat3

1 2

Summary of Contents

Page 1 - (Mammalian Derived)

#5358SterileHuman Interleukin-10 (hIL-10)(Mammalian Derived)New 07/10n SC 10 µg (With Carrier)n LC 50 µg (With Carrier)n SF 10 µg (Carrier Free)

Page 2 - (mammalian derived)

Orders n 877-616-CELL (2355) [email protected] Support n 877-678-TECH (8324) [email protected] Web n www.cellsignal.co

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Summary of Contents

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