Invitrogen

IL-4 Monoclonal Antibody (25D2)

Tech Support

Datasheet

Protocols

Questions & Answers

Tech Support

Product Image Gallery

IL-4 Antibody in Western Blot (WB) — 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).

IL-4 Antibody in ELISA (ELISA) — 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.

Product Details

M450

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000	View 2 publications 2 publications
ELISA (ELISA)	0.006-25 µg/mL	View 8 publications 8 publications
Immunohistochemistry (IHC)	-	View 2 publications 2 publications
Miscellaneous PubMed (Misc)	-	View 2 publications 2 publications
Flow Cytometry (Flow)	-	View 9 publications 9 publications
Immunocytochemistry (ICC/IF)	-	View 1 publication 1 publication
Neutralization (Neu)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human
Published species	Human, Mouse
Host/Isotype	Rat / IgG1
Class	Monoclonal
Type	Antibody
Clone	25D2
Immunogen	Recombinant human IL-4 (CHO cell-derived)
Conjugate	Unconjugated Biotin
Form	Liquid
Concentration	1.0 mg/mL
Purification	Protein G
Storage buffer	PBS
Contains	no preservative
Storage conditions	-20°C
RRID	AB_223568

Product Specific Information

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).

Target Information

Interleukin-4 (IL-4) is a pleiotropic, immune-modulatory cytokine produced by activated T cells, and serves as a ligand for interleukin 4 receptor. IL-4 has been found to have the broadest range of actions. The interleukin 4 receptor also binds to IL13, which may contribute to many overlapping functions of this cytokine and IL13. STAT6, a signal transducer and activator of transcription, has been shown to play an important role in mediating the immune regulatory signal of IL-4. The IL-4 gene, IL-3, IL-5, IL-13, and CSF2 form a cytokine gene cluster on chromosome 5q. In particular, IL-4 is located in close proximity to IL13 on the chromosome. Further, the IL-4 gene, IL13 and IL5 are found to be coordinated by several long-range regulatory elements over a 120 kilobase range on the chromosome. IL-4 delta 2 is a known splice variant of IL-4.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

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

Biotechnology and bioengineering

Youngblood R,Flesher CG,Delproposto J,Baker NA,Neeley CK,Li F,Lumeng CN,Shea LD,O'Rourke RW

M450 was used in Immunohistochemistry to demonstrate that polymer biomaterials implanted into adipose tissue have the potential to modulate local tissue and systemic inflammation and metabolism.

Tue Dec 01 00:00:00 EST 2020

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

Journal of immunology (Baltimore, Md. : 1950)

Sung SJ,Ge Y,Dai C,Wang H,Fu SM,Sharma R,Hahn YS,Yu J,Le TH,Okusa MD,Bolton WK,Lawler JR

M450 was used in Magnetic cell separation to identify glomerulus-infiltrating leukocyte populations in NZM2328 lupus-prone mice with spontaneous chronic glomerulonephritis and anti-glomerular basement membrane-induced nephritis.

Sat Apr 01 00:00:00 EDT 2017

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

PloS one

Janardhan SV,Marks R,Gajewski TF

M450 was used in Western Blotting to demonstrate that constitutive Ras signaling inhibits the ability of CD4+ T-cells to properly differentiate into Th1/Th2 effector cytokine-producing cells.

Wed Dec 30 00:00:00 EST 2015

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

The Journal of experimental medicine

Tesio M,Tang Y,Müdder K,Saini M,von Paleske L,Macintyre E,Pasparakis M,Waisman A,Trumpp A

M450 was used in Flow cytometry/Cell sorting to elucidate the molecular mechanisms regulating haematopoietic stem cell dormancy.

Mon Apr 06 00:00:00 EDT 2015

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

PloS one

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,Chen YT,Chen CH,Wu JY,Lee YC

M450 was used in Flow cytometry/Cell sorting to suggest that peripheral B cells play a pathogenic role during the acute stage of Kawasaki disease.

Tue Apr 15 00:00:00 EDT 2014

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

The Journal of experimental medicine

Tesio M,Oser GM,Baccelli I,Blanco-Bose W,Wu H,Göthert JR,Kogan SC,Trumpp A

M450 was used in Magnetic cell separation to investigate the effect of PTEN deletion in bone marrow, showing that it leads to G-CSF-mediated hematopoietic stem cell mobilisation.

Mon Oct 21 00:00:00 EDT 2013

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

PloS one

Wolff MJ,Leung JM,Davenport M,Poles MA,Cho I,Loke P

M450 was used in flow cytometry to determine the localization of CD4 positive T cells subsets in the healthy intestine

Fri Dec 07 00:00:00 EST 2012

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

Pathology international

Ali A,Furusato B,Ts'o PO,Lum ZP,Elsamanoudi S,Mohamed A,Srivastava S,Moul JW,Brassell SA,Sesterhenn IA,McLeod DG

M450 was used in Flow cytometry/Cell sorting to assess the incidence of circulating tumor cells (CTCs) in prostate cancer patients with low-volume tumors (less than 0.5 cc) after radical prostatectomy (RP).

Fri Oct 01 00:00:00 EDT 2010

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

Biochemical and biophysical research communications

Curbo S,Gaudin R,Carlsten M,Malmberg KJ,Troye-Blomberg M,Ahlborg N,Karlsson A,Johansson M,Lundberg M

M450 was used in ELISA to study the effect of disulfide bonds on interleukin-4 signaling

Fri Dec 25 00:00:00 EST 2009

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

Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology

Nilsson C,Larsson Sigfrinius AK,Montgomery SM,Sverremark-Ekström E,Linde A,Lilja G,Blomberg MT

M450 was used in ELISA to investigate the relationship between the cytokine profile in PBMC and Epstein-Barr virus serostatus and between the cytokine profile and cytomegalovirus serostatus

Wed Apr 01 00:00:00 EDT 2009

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

Biophysical journal

Robert P,Sengupta K,Puech PH,Bongrand P,Limozin L

M450 was used in Flow cytometry/Cell sorting to open the way for further quantitative studies of the effect of molecular environment and separation distance on ligand-receptor association and dissociation.

Wed Oct 01 00:00:00 EDT 2008

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

Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver

Marotti B,Rocco A,De Colibus P,Compare D,de Nucci G,Staibano S,Tatangelo F,Romano M,Nardone G

M450 was used in ELISA to study the mucosal production of interleukin 13 in Helicobacter pylori-related gastric diseases

Tue Apr 01 00:00:00 EDT 2008

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

Journal of virology

Kueng HJ,Leb VM,Haiderer D,Raposo G,Thery C,Derdak SV,Schmetterer KG,Neunkirchner A,Sillaber C,Seed B,Pickl WF

M450 was used in immunocytochemistry to develop a system for the decoration of enveloped viruses with functionally active cytokines

Wed Aug 01 00:00:00 EDT 2007

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

European journal of immunology

Bratke K,Kuepper M,Bade B,Virchow JC,Luttmann W

M450 was used in flow cytometry to use flow cytometry to study granzyme A, B, and K expression in human peripheral blood lymphocytes

Thu Sep 01 00:00:00 EDT 2005

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

Respiratory research

Chialda L,Zhang M,Brune K,Pahl A

Published figure using IL-4 monoclonal antibody (Product # M450) in ELISA

Fri Apr 15 00:00:00 EDT 2005

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

Clinical and experimental immunology

Melgar S,Yeung MM,Bas A,Forsberg G,Suhr O,Oberg A,Hammarstrom S,Danielsson A,Hammarstrom ML

M450 was used in immunohistochemistry to study the involvement of interleukin 10 in active ulcerative colitis

Wed Oct 01 00:00:00 EDT 2003

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

The Journal of rheumatology

Hatachi S,Iwai Y,Kawano S,Morinobu S,Kobayashi M,Koshiba M,Saura R,Kurosaka M,Honjo T,Kumagai S

M450 was used in flow cytometry to investigate the involvement of the PD-1 system in rheumatoid arthritis

Tue Jul 01 00:00:00 EDT 2003

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

Journal of hepatology

Loffreda S,Muratori P,Muratori L,Mele L,Bianchi FB,Lenzi M

M450 was used in ELISA to investigate the augmented production of Th1-type cytokines in HCV-infected cryoglobulinemic patients

Sat Feb 01 00:00:00 EST 2003

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

Journal of immunology (Baltimore, Md. : 1950)

Dimitrova P,Skapenko A,Herrmann ML,Schleyerbach R,Kalden JR,Schulze-Koops H

M450 was used in blocking/activating experiment to investigate the role of de novo pyrimidine biosynthesis restriction for Th1 cell activation and Th2 cell differentiation

Sun Sep 15 00:00:00 EDT 2002

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

Clinical and experimental immunology

Gabrielsson S,Söderlund A,Nilsson C,Lilja G,Nordlund M,Troye-Blomberg M

M450 was used in ELISA to study the effect of fetal environment on the development of atopic disease

Sat Dec 01 00:00:00 EST 2001

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

Journal of immunology (Baltimore, Md. : 1950)

Skapenko A,Lipsky PE,Kraetsch HG,Kalden JR,Schulze-Koops H

M450 was used in ELISA to investigate the role of interleukin 4 gene expression

Sun Apr 01 00:00:00 EST 2001

Functional expression of transmembrane 4 superfamily molecules on human eosinophils.

International archives of allergy and immunology

Matsumoto K,Bochner BS,Wakiguchi H,Kurashige T

M450 was used in Flow cytometry/Cell sorting to suggest that several TM4SF molecules are functionally expressed on human eosinophils, and therefore might participate in allergic inflammation.

Tue Nov 09 00:00:00 EST 1999

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

Molecular and cellular biology

Ossipow V,Fonjallaz P,Schibler U

M450 was used in Western Blotting to study the interaction between an RNA polymerase II complex and thyroid embryonic factor.

Mon Feb 01 00:00:00 EST 1999

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

International archives of allergy and immunology

Matsumoto K,Bochner BS,Wakiguchi H,Kurashige T

M450 was used in Flow cytometry/Cell sorting to suggest that CD45 isoforms are functionally expressed on human eosinophils, and are capable of modulating eosinophil function and might participate in allergic inflammation.

Tue Sep 01 00:00:00 EDT 1998

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

The Journal of experimental medicine

Anstey NM,Weinberg JB,Hassanali MY,Mwaikambo ED,Manyenga D,Misukonis MA,Arnelle DR,Hollis D,McDonald MI,Granger DL

M450 was used in ELISA to measure the interleukin-10 and interleukin-4 level in Tanzanian children with malaria

Thu Aug 01 00:00:00 EDT 1996

Bioinformatics

Protein Aliases: B cell growth factor 1; B-cell stimulatory factor 1; B_cell stimulatory factor 1; Binetrakin; BSF-1; H-IL-4; IL-4; ILN; Interleukin; interleukin 4 variant 2; Interleukin-4; Interleukin4; Lymphocyte stimulatory factor 1; M-IL-4; MGC79402; Pitrakinra

Gene Aliases: BCGF-1; BCGF1; BSF-1; BSF1; IL-4; IL4

UniProt ID: (Human) P05112

Entrez Gene ID: (Human) 3565

Molecular Function: interleukin superfamily

Function(s)

Process(es)

cytokine activity interleukin-4 receptor binding protein binding growth factor activity

IL-4 Monoclonal Antibody (25D2)

Product Details

M450

Western Blot (WB)

ELISA (ELISA)

Immunohistochemistry (IHC)

Miscellaneous PubMed (Misc)

Flow Cytometry (Flow)

Immunocytochemistry (ICC/IF)

Neutralization (Neu)

Species Reactivity

Published species

Host/Isotype

Class

Type

Clone

Immunogen

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

Performance Guarantee

We're here to help

Your items have has been added!