M450

antibody from Invitrogen Antibodies

Targeting: IL4 BCGF-1, BCGF1, BSF1, IL-4, MGC79402

Provider product page for M450

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Antibody data

Antibody Data
Antigen structure
References [31]
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Validations
Western blot [1]
Other assay [2]

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Product number: M450 - Provider product page
Provider: Invitrogen Antibodies
Product name: IL-4 Monoclonal Antibody (25D2)
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: M450 targets IL-4 in WB and ELISA applications and shows reactivity with Human samples. The M450 immunogen is recombinant human IL-4 (CHO cell-derived). M450 detects IL-4 which has a predicted molecular weight of approximately 15 kDa. 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: 25D2
Vial size: 500 µg
Concentration: 1.0 mg/mL
Storage: -20°C

IL4 protein structure - M450 shown in red.

Submitted references Regulation of adipose tissue inflammation and systemic metabolism in murine obesity by polymer implants loaded with lentiviral vectors encoding human interleukin-4.

Youngblood R, Flesher CG, Delproposto J, Baker NA, Neeley CK, Li F, Lumeng CN, Shea LD, O'Rourke RW
Biotechnology and bioengineering 2020 Dec;117(12):3891-3901

Dependence of Glomerulonephritis Induction on Novel Intraglomerular Alternatively Activated Bone Marrow-Derived Macrophages and Mac-1 and PD-L1 in Lupus-Prone NZM2328 Mice.

Sung SJ, Ge Y, Dai C, Wang H, Fu SM, Sharma R, Hahn YS, Yu J, Le TH, Okusa MD, Bolton WK, Lawler JR
Journal of immunology (Baltimore, Md. : 1950) 2017 Apr 1;198(7):2589-2601

Hematopoietic stem cell quiescence and function are controlled by the CYLD-TRAF2-p38MAPK pathway.

Tesio M, Tang Y, Müdder K, Saini M, von Paleske L, Macintyre E, Pasparakis M, Waisman A, Trumpp A
The Journal of experimental medicine 2015 Apr 6;212(4):525-38

Primary murine CD4+ T cells fail to acquire the ability to produce effector cytokines when active Ras is present during Th1/Th2 differentiation.

Janardhan SV, Marks R, Gajewski TF
PloS one 2014;9(11):e112831

Replication and meta-analysis of GWAS identified susceptibility loci in Kawasaki disease confirm the importance of B lymphoid tyrosine kinase (BLK) in disease susceptibility.

Chang CJ, Kuo HC, Chang JS, Lee JK, Tsai FJ, Khor CC, Chang LC, Chen SP, Ko TM, Liu YM, Chen YJ, Hong YM, Jang GY, Hibberd ML, Kuijpers T, Burgner D, Levin M, Burns JC, Davila S, International Kawasaki Disease Genetics Consortium, Korean Kawasaki Disease Genetics Consortium, Taiwan Kawasaki Disease Genetics Consortium, Chen YT, Chen CH, Wu JY, Lee YC
PloS one 2013;8(8):e72037

Pten loss in the bone marrow leads to G-CSF-mediated HSC mobilization.

Tesio M, Oser GM, Baccelli I, Blanco-Bose W, Wu H, Göthert JR, Kogan SC, Trumpp A
The Journal of experimental medicine 2013 Oct 21;210(11):2337-49

TH17, TH22 and Treg cells are enriched in the healthy human cecum.

Wolff MJ, Leung JM, Davenport M, Poles MA, Cho I, Loke P
PloS one 2012;7(7):e41373

Assessment of circulating tumor cells (CTCs) in prostate cancer patients with low-volume tumors.

Ali A, Furusato B, Ts'o PO, Lum ZP, Elsamanoudi S, Mohamed A, Srivastava S, Moul JW, Brassell SA, Sesterhenn IA, McLeod DG
Pathology international 2010 Oct;60(10):667-72

Regulation of interleukin-4 signaling by extracellular reduction of intramolecular disulfides.

Curbo S, Gaudin R, Carlsten M, Malmberg KJ, Troye-Blomberg M, Ahlborg N, Karlsson A, Johansson M, Lundberg M
Biochemical and biophysical research communications 2009 Dec 25;390(4):1272-7

Epstein-Barr virus and cytomegalovirus are differentially associated with numbers of cytokine-producing cells and early atopy.

Nilsson C, Larsson Sigfrinius AK, Montgomery SM, Sverremark-Ekström E, Linde A, Lilja G, Blomberg MT
Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology 2009 Apr;39(4):509-17

Tuning the formation and rupture of single ligand-receptor bonds by hyaluronan-induced repulsion.

Robert P, Sengupta K, Puech PH, Bongrand P, Limozin L
Biophysical journal 2008 Oct;95(8):3999-4012

Interleukin-13 mucosal production in Helicobacter pylori-related gastric diseases.

Marotti B, Rocco A, De Colibus P, Compare D, de Nucci G, Staibano S, Tatangelo F, Romano M, Nardone G
Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver 2008 Apr;40(4):240-7

General strategy for decoration of enveloped viruses with functionally active lipid-modified cytokines.

Kueng HJ, Leb VM, Haiderer D, Raposo G, Thery C, Derdak SV, Schmetterer KG, Neunkirchner A, Sillaber C, Seed B, Pickl WF
Journal of virology 2007 Aug;81(16):8666-76

Differential expression of human granzymes A, B, and K in natural killer cells and during CD8+ T cell differentiation in peripheral blood.

Bratke K, Kuepper M, Bade B, Virchow JC Jr, Luttmann W
European journal of immunology 2005 Sep;35(9):2608-16

Human rhinoviruses inhibit the accessory function of dendritic cells by inducing sialoadhesin and B7-H1 expression.

Kirchberger S, Majdic O, Steinberger P, Blüml S, Pfistershammer K, Zlabinger G, Deszcz L, Kuechler E, Knapp W, Stöckl J
Journal of immunology (Baltimore, Md. : 1950) 2005 Jul 15;175(2):1145-52

Inhibitors of mitogen-activated protein kinases differentially regulate costimulated T cell cytokine production and mouse airway eosinophilia.

Chialda L, Zhang M, Brune K, Pahl A
Respiratory research 2005 Apr 15;6(1):36

Over-expression of interleukin 10 in mucosal T cells of patients with active ulcerative colitis.

Melgar S, Yeung MM, Bas A, Forsberg G, Suhr O, Oberg A, Hammarstrom S, Danielsson A, Hammarstrom ML
Clinical and experimental immunology 2003 Oct;134(1):127-37

Over-expression of interleukin 10 in mucosal T cells of patients with active ulcerative colitis.

Melgar S, Yeung MM, Bas A, Forsberg G, Suhr O, Oberg A, Hammarstrom S, Danielsson A, Hammarstrom ML
Clinical and experimental immunology 2003 Oct;134(1):127-37

CD4+ PD-1+ T cells accumulate as unique anergic cells in rheumatoid arthritis synovial fluid.

Hatachi S, Iwai Y, Kawano S, Morinobu S, Kobayashi M, Koshiba M, Saura R, Kurosaka M, Honjo T, Kumagai S
The Journal of rheumatology 2003 Jul;30(7):1410-9

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

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

Restriction of de novo pyrimidine biosynthesis inhibits Th1 cell activation and promotes Th2 cell differentiation.

Dimitrova P, Skapenko A, Herrmann ML, Schleyerbach R, Kalden JR, Schulze-Koops H
Journal of immunology (Baltimore, Md. : 1950) 2002 Sep 15;169(6):3392-9

Influence of atopic heredity on IL-4-, IL-12- and IFN-gamma-producing cells in in vitro activated cord blood mononuclear cells.

Gabrielsson S, Söderlund A, Nilsson C, Lilja G, Nordlund M, Troye-Blomberg M
Clinical and experimental immunology 2001 Dec;126(3):390-6

Influence of atopic heredity on IL-4-, IL-12- and IFN-gamma-producing cells in in vitro activated cord blood mononuclear cells.

Gabrielsson S, Söderlund A, Nilsson C, Lilja G, Nordlund M, Troye-Blomberg M
Clinical and experimental immunology 2001 Dec;126(3):390-6

Antigen-independent Th2 cell differentiation by stimulation of CD28: regulation via IL-4 gene expression and mitogen-activated protein kinase activation.

Skapenko A, Lipsky PE, Kraetsch HG, Kalden JR, Schulze-Koops H
Journal of immunology (Baltimore, Md. : 1950) 2001 Apr 1;166(7):4283-92

Antigen-independent Th2 cell differentiation by stimulation of CD28: regulation via IL-4 gene expression and mitogen-activated protein kinase activation.

Skapenko A, Lipsky PE, Kraetsch HG, Kalden JR, Schulze-Koops H
Journal of immunology (Baltimore, Md. : 1950) 2001 Apr 1;166(7):4283-92

Functional expression of transmembrane 4 superfamily molecules on human eosinophils.

Matsumoto K, Bochner BS, Wakiguchi H, Kurashige T
International archives of allergy and immunology 1999;120 Suppl 1:38-44

An RNA polymerase II complex containing all essential initiation factors binds to the activation domain of PAR leucine zipper transcription factor thyroid embryonic factor.

Ossipow V, Fonjallaz P, Schibler U
Molecular and cellular biology 1999 Feb;19(2):1242-50

Altered expression of CD11b and CD62L after cross-linking of CD45 isoforms on human eosinophils.

Matsumoto K, Bochner BS, Wakiguchi H, Kurashige T
International archives of allergy and immunology 1998 Sep;117 Suppl 1:34-9

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

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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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of IL-4 was performed by loading 1 µg of human recombiannt protein IL-4 (1 µg/lane, Product # RIL4I) in non-reducing sample buffer and 8 µL PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BOX). Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288) and blocked with 5% Milk/TBST for at least 1 hour at room temperature. IL-4 was detected using a IL-4 rat monoclonal antibody (Product # M450) at a dilution of 1:1000 in blocking buffer overnight at 4°C on a rocking platform, followed by a HRP conjugated secondary antibody (Product # 62-9520) at a dilution of 1:5000 for at least 1 hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Dura Extended Duration Substrate (Product # 34076) and the myECL Imager (Product # 62236).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Direct ELISA analysis of IL-4 was performed by coating wells of a 96-well plate with 100 µL per well of IL-4 (Product # RIL4I) diluted in carbonate/bicarbonate buffer (Product # 28382) at a concentration of 1 µg/mL overnight at 4C. Wells of the plate were washed, blocked with starting blocking buffer (Product # 37538), and incubated with 100 µL per well of a rat anti-IL-4 monoclonal antibody (Product # M450) at a concentration of 0.006, 0.024, 0.097, 0.390, 1.560, 6.250 and 25 µg/mL for 90 minutes at 37C. The plate was washed, then incubated with 100 µL per well of an HRP-conjugated goat anti-rat IgG secondary antibody (Product # 62-9520) at a dilution of 1:1000 for 90 minutes at 37C. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 5 minutes at room temperature in the dark. The reaction was stopped with Stop solution (Product # N600), and absorbances were read on a spectrophotometer at 450-550 nm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Induction of cytokines in CD4+ T cells by different costimulatory receptors. Human CD4 + T cells were stimulated with various stimuli. (A) Cytokine mRNA level were determined after eight hours stimulation using real-time RT-PCR. mRNA level were normalized to beta-actin and CD3 stimulated cells were set to 1. Each column represents mean +- s.e.mean of three different donors. (B) Cytokine protein level was determined in the supernatant by ELISA after twenty-four hours of stimulation. Each column represents mean +- s.e.mean of three different volunteers. *P < 0.05 (versus CD3 stimulated samples).