M010

antibody from Invitrogen Antibodies

Targeting: IL10 CSIF, IL-10, IL10A, TGIF

Western blot (Supportive data in Antibodypedia)

ELISA (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

M010

Provider

Invitrogen Antibodies

Product name

IL-10 Monoclonal Antibody (9D7)

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

M010 targets IL-10 in ELISA, and IA applications and shows reactivity with Human samples. The M010 immunogen is recombinant human IL-10 (COS cell-derived). M010 detects IL-10 which has a predicted molecular weight of approximately 19 kDa. The M010 IL10 antibody (clone 9D7) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M011B (biotinylated conjugate of clone 12G8). Typical dilutions for sandwich ELISA include 1 µg/mL for coating and 0.125 - 0.25 µg/mL for detection. Antibody M010 (clone 9D7) and biotinylated antibody M011B (clone 12G8) have successfully been used in combination with recombinant IL10 protein SIL10 in ELISA applications. This antibody is produced by injecting Rat IgG secreting hybridoma cells into the peritoneum of mice. The resulting ascites is collected from the mice and the antibody is purified. This product has been tested for endotoxins by limulus amoebocyte lysate (LAL) assay and contains an endotoxin concentration of less than or equal to 10 endotoxin units per milligram (EU/mg).

Reactivity

Human

Host

Rat

Isotype

IgG

Antibody clone number

9D7

Vial size

500 µg

Concentration

1.0 mg/mL

Storage

-20°C

References

Submitted references

Methods for in vitro generation of human type 1 regulatory T cells.

Gregori S, Roncarolo MG, Bacchetta R
Methods in molecular biology (Clifton, N.J.) 2011;677:31-46

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Hepatitis C virus modulates human monocyte-derived dendritic cells.

Eksioglu EA, Bess JR, Zhu H, Xu Y, Dong HJ, Elyar J, Nelson DR, Liu C
Journal of viral hepatitis 2010 Nov;17(11):757-69

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Molecular and functional characterization of allogantigen-specific anergic T cells suitable for cell therapy.

Bacchetta R, Gregori S, Serafini G, Sartirana C, Schulz U, Zino E, Tomiuk S, Jansen U, Ponzoni M, Paties CT, Fleischhauer K, Roncarolo MG
Haematologica 2010 Dec;95(12):2134-43

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Type 1 regulatory T cells are associated with persistent split erythroid/lymphoid chimerism after allogeneic hematopoietic stem cell transplantation for thalassemia.

Serafini G, Andreani M, Testi M, Battarra M, Bontadini A, Biral E, Fleischhauer K, Marktel S, Lucarelli G, Roncarolo MG, Bacchetta R
Haematologica 2009 Oct;94(10):1415-26

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Anti-ribosomal phosphoprotein autoantibody triggers interleukin-10 overproduction via phosphatidylinositol 3-kinase-dependent signalling pathways in lipopolysaccharide-activated macrophages.

Lee TP, Leu SJ, Huang JC, Song YC, Jhou RS, Tang SJ, Sun KH
Immunology 2009 May;127(1):91-102

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Multiplexed Elispot assay.

Harriman WD, Collarini EJ, Cromer RG, Dutta A, Strandh M, Zhang F, Kauvar LM
Journal of immunological methods 2009 Feb 28;341(1-2):127-34

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Latent membrane protein 1 of EBV activates phosphatidylinositol 3-kinase to induce production of IL-10.

Lambert SL, Martinez OM
Journal of immunology (Baltimore, Md. : 1950) 2007 Dec 15;179(12):8225-34

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Hepatitis C virus-specific reactivity of CD4+-lymphocytes in children born from HCV-infected women.

Della Bella S, Riva A, Tanzi E, Nicola S, Amendola A, Vecchi L, Nebbia G, Longhi R, Zanetti AR, Villa ML
Journal of hepatology 2005 Sep;43(3):394-402

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Modulation of human dendritic cell phenotype and function by probiotic bacteria.

Hart AL, Lammers K, Brigidi P, Vitali B, Rizzello F, Gionchetti P, Campieri M, Kamm MA, Knight SC, Stagg AJ
Gut 2004 Nov;53(11):1602-9

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Generation of three-dimensional pannus-like tissues in vitro from single cell suspensions of synovial fluid cells from arthritis patients.

Solomon S, Masilamani M, Mohanty S, Schwab JE, Boneberg EM, Illges H
Rheumatology international 2004 Mar;24(2):71-6

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Host factors impacting the innate response in humans to the candidate adjuvants RC529 and monophosphoryl lipid A.

Tiberio L, Fletcher L, Eldridge JH, Duncan DD
Vaccine 2004 Mar 29;22(11-12):1515-23

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Long-term treatment with etanercept significantly reduces the number of proinflammatory cytokine-secreting peripheral blood mononuclear cells in patients with rheumatoid arthritis.

Schotte H, Schlüter B, Willeke P, Mickholz E, Schorat MA, Domschke W, Gaubitz M
Rheumatology (Oxford, England) 2004 Aug;43(8):960-4

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Measles virus infection of rhesus macaques affects neutrophil expression of IL-12 and IL-10.

Hoffman SJ, Polack FP, Hauer DA, Griffin DE
Viral immunology 2003;16(3):369-79

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Altered maturation of peripheral blood dendritic cells in patients with breast cancer.

Della Bella S, Gennaro M, Vaccari M, Ferraris C, Nicola S, Riva A, Clerici M, Greco M, Villa ML
British journal of cancer 2003 Oct 20;89(8):1463-72

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Monocyte chemoattractant protein-1 levels in bronchoalveolar lavage fluid of lung-transplanted patients treated with tacrolimus as rescue treatment for refractory acute rejection.

Meloni F, Cascina A, Paschetto E, Marone Bianco A, Morosini M, Pellegrini C, Fietta A, Vitulo P, Pozzi E, Viganò M
Transplantation proceedings 2003 Jun;35(4):1523-6

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Monocyte chemoattractant protein-1 levels in bronchoalveolar lavage fluid of lung-transplanted patients treated with tacrolimus as rescue treatment for refractory acute rejection.

Meloni F, Cascina A, Paschetto E, Marone Bianco A, Morosini M, Pellegrini C, Fietta A, Vitulo P, Pozzi E, Viganò M
Transplantation proceedings 2003 Jun;35(4):1523-6

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Enhanced monocyte Th1 cytokine production in HCV-infected cryoglobulinemic patients.

Loffreda S, Muratori P, Muratori L, Mele L, Bianchi FB, Lenzi M
Journal of hepatology 2003 Feb;38(2):230-6

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Enhanced monocyte Th1 cytokine production in HCV-infected cryoglobulinemic patients.

Loffreda S, Muratori P, Muratori L, Mele L, Bianchi FB, Lenzi M
Journal of hepatology 2003 Feb;38(2):230-6

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The neuropeptide bombesin improves IgA-mediated mucosal immunity with preservation of gut interleukin-4 in total parenteral nutrition-fed mice.

Zarzaur BL, Wu Y, Fukatsu K, Johnson CD, Kudsk KA
Surgery 2002 Jan;131(1):59-65

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The neuropeptide bombesin improves IgA-mediated mucosal immunity with preservation of gut interleukin-4 in total parenteral nutrition-fed mice.

Zarzaur BL, Wu Y, Fukatsu K, Johnson CD, Kudsk KA
Surgery 2002 Jan;131(1):59-65

---

Ambient air particulates stimulate alveolar macrophages of smokers to promote differentiation of myeloid precursor cells.

Suwa T, Hogg JC, Vincent R, Mukae H, Fujii T, van Eeden SF
Experimental lung research 2002 Jan-Feb;28(1):1-18

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Ambient air particulates stimulate alveolar macrophages of smokers to promote differentiation of myeloid precursor cells.

Suwa T, Hogg JC, Vincent R, Mukae H, Fujii T, van Eeden SF
Experimental lung research 2002 Jan-Feb;28(1):1-18

---

Altered synthesis of interleukin-12 and type 1 and type 2 cytokinesin rhesus macaques during measles and atypical measles.

Polack FP, Hoffman SJ, Moss WJ, Griffin DE
The Journal of infectious diseases 2002 Jan 1;185(1):13-9

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Leishmania priming of human dendritic cells for CD40 ligand-induced interleukin-12p70 secretion is strain and species dependent.

McDowell MA, Marovich M, Lira R, Braun M, Sacks D
Infection and immunity 2002 Aug;70(8):3994-4001

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Leishmania priming of human dendritic cells for CD40 ligand-induced interleukin-12p70 secretion is strain and species dependent.

McDowell MA, Marovich M, Lira R, Braun M, Sacks D
Infection and immunity 2002 Aug;70(8):3994-4001

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Adaptive immune responses of patients with asthma to the attachment (G) glycoprotein of respiratory synctial virus.

Hancock GE, Scheuer CA, Sierzega R, Pryharski KS, McBride JT, Watelet LF, Tebbey PW, Smith JD
The Journal of infectious diseases 2001 Dec 15;184(12):1589-93

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Autoregulation of human monocyte-derived dendritic cell maturation and IL-12 production by cyclooxygenase-2-mediated prostanoid production.

Whittaker DS, Bahjat KS, Moldawer LL, Clare-Salzler MJ
Journal of immunology (Baltimore, Md. : 1950) 2000 Oct 15;165(8):4298-304

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Autoregulation of human monocyte-derived dendritic cell maturation and IL-12 production by cyclooxygenase-2-mediated prostanoid production.

Whittaker DS, Bahjat KS, Moldawer LL, Clare-Salzler MJ
Journal of immunology (Baltimore, Md. : 1950) 2000 Oct 15;165(8):4298-304

---

Donor-specific stimulation of peripheral blood mononuclear cells from recipients of orthotopic liver transplants is associated, in the absence of rejection, with type-2 cytokine production.

Chung SW, Yoshida EM, Cattral MS, Hu Y, Gorczynski RM
Immunology letters 1998 Sep;63(2):91-6

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Nitric oxide in Tanzanian children with malaria: inverse relationship between malaria severity and nitric oxide production/nitric oxide synthase type 2 expression.

Anstey NM, Weinberg JB, Hassanali MY, Mwaikambo ED, Manyenga D, Misukonis MA, Arnelle DR, Hollis D, McDonald MI, Granger DL
The Journal of experimental medicine 1996 Aug 1;184(2):557-67

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Nitric oxide in Tanzanian children with malaria: inverse relationship between malaria severity and nitric oxide production/nitric oxide synthase type 2 expression.

Anstey NM, Weinberg JB, Hassanali MY, Mwaikambo ED, Manyenga D, Misukonis MA, Arnelle DR, Hollis D, McDonald MI, Granger DL
The Journal of experimental medicine 1996 Aug 1;184(2):557-67

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Contribution of IL-1, CD14, and CD13 in the increased IL-6 production induced by in vitro monocyte-synoviocyte interactions.

Chomarat P, Rissoan MC, Pin JJ, Banchereau J, Miossec P
Journal of immunology (Baltimore, Md. : 1950) 1995 Oct 1;155(7):3645-52

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-IL-10 Monoclonal Antibody (9D7) (Product # M010) and a 18kDa band corresponding to Interleukin-10 was observed in Human rec. IL10 as compared to Human rec. IL6 which was used as a negative control. Mouse rec. IL6 was used to test species reactivity. 250 ng of Human rec. IL10 (Lane 1), Human rec. IL6 (Lane 2), Mouse rec. IL10 (Lane 3) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with F(ab'2-Rabbit anti-Rat IgG (H+L Secondary Antibody, HRP (Product # PA1-29927, 1:20000 dilution) using the iBright FL 1500 (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).

ELISA

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Sandwich ELISA analysis of human IL-10 was performed using a Human IL-10 Colorimetric ELISA kit (Product # EHIL10) by loading 50 µL per well of Human IL-10 recombinant protein (Product # SIL10) in triplicate at 600, 240, 96, 38.4, 15.36, and 0 pg/mL across a 2 µg/mL rat anti-human IL-10 (Product # M010) pre-coated plate and incubating for 1.5 hours at room temperature. The plate was washed, and incubated with 50 µL per well of an IL-10 biotinylated antibody (Product # M011B) in triplicate at 0.3 µg/mL for 1.5 hours in conjunction with standards at room temperature. The plate was washed and incubated with 100 µL per well of Ultra Streptavidin-HRP (Product # N504) in all test wells at a dilution of 1:18,000 for 15 minutes at room temperature. Detection was performed using TMB Substrate Solution (Product # N301) for 20 minutes at room temperature in the dark. The reaction was then stopped with 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Sandwich ELISA analysis of human IL-10 was performed using a Human IL-10 Colorimetric ELISA kit (Product # EHIL10) by loading 50 µL per well of Human IL-10 recombinant protein (Product # SIL10) in triplicate at 600, 240, 96, 38.4, 15.36, and 0 pg/mL across a 2 µg/mL rat anti-human IL-10 (Product # M010) pre-coated plate and incubating for 1.5 hours at room temperature. The plate was washed, and incubated with 50 µL per well of an IL-10 biotinylated antibody (Product # M011B) in triplicate at 0.3 µg/mL for 1.5 hours in conjunction with standards at room temperature. The plate was washed and incubated with 100 µL per well of Ultra Streptavidin-HRP (Product # N504) in all test wells at a dilution of 1:18,000 for 15 minutes at room temperature. Detection was performed using TMB Substrate Solution (Product # N301) for 20 minutes at room temperature in the dark. The reaction was then stopped with 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.