M450B

antibody from Invitrogen Antibodies

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

Western blot (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

M450B

Provider

Invitrogen Antibodies

Product name

IL-4 Monoclonal Antibody (25D2), Biotin

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

M450B targets IL-4 in WB and ELISA applications and shows reactivity with Human samples.

Conjugate

Biotin

Antibody clone number

25D2

Concentration

0.5 mg/mL

References

Submitted references

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

---

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

---

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

Western blot

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 biotin conjugated IL-4 rat monoclonal antibody (Product # M450B) at a dilution of 1:1000 in blocking buffer overnight at 4°C on a rocking platform, followed by a pre-diluted High Sensitivity Streptavidin-HRP antibody (Product # 21134) at a dilution of 1:50 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).

Conjugate

Biotin

Other assay

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 biotin conjugated anti-IL-4 monoclonal antibody (Product # M450B) at a concentration of 0.003, 0.012, 0.049, 0.195, 0.780, 3.125 and 12.50 µg/mL for 90 minutes at 37C. The plate was washed, then incubated with 100 µL per well of pre-diluted High Sensitivity Streptavidin-HRP antibody (Product # 21134) at a dilution of 1:50 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.

Conjugate

Biotin