M3IL4I

antibody from Invitrogen Antibodies

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

Western blot (Supportive data in Antibodypedia)

ELISA (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

M3IL4I

Provider

Invitrogen Antibodies

Product name

IL-4 Monoclonal Antibody (E10023)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

M3IL4I targets IL-4 in ELISA and Western blot applications and shows reactivity with Human samples. The M3IL4I immunogen is recombinant human IL-4 purified from Pichia pastoris. M3IL4I detects IL-4 which has a predicted molecular weight of approximately 15 kDa. The M3IL4I IL4 antibody (clone E10023) has successfully been paired as the coating antibody in a sandwich ELISA with detection antibody M2IL4BI (biotinylated conjugate of clone E10022). Typical dilutions for sandwich ELISA include 1 µg/mL for coating and 0.125 - 0.25 µg/mL for detection. Antibody M3IL4I (clone E10022) and biotinylated antibody M2IL4BI (clone E10023) have successfully been used in combination with recombinant IL4 protein SIL4 in ELISA applications. Endotoxin levels are less than or equal to 10 µg/mL.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

E10023

Vial size

500 µg

Concentration

0.5 mg/mL

Storage

4° C

References

Submitted references

De-immunized and Functional Therapeutic (DeFT) versions of a long lasting recombinant alpha interferon for antiviral therapy.

Mufarrege EF, Giorgetti S, Etcheverrigaray M, Terry F, Martin W, De Groot AS
Clinical immunology (Orlando, Fla.) 2017 Mar;176:31-41

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 IL-4 mouse monoclonal antibody (Product # M3IL4I) 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 # 62-6520) 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).

ELISA

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 mouse anti-IL-4 monoclonal antibody (Product # M3IL4I) at a concentration of 0.005, 0.020, 0.078, 0.312, 1.25, 5 and 20 µg/mL for 90 minutes at 37C. The plate was washed, then incubated with 100 µL per well of an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 62-6520) at a dilution of 1:2000 for 90 minutes at 37C. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 30 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.

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 mouse anti-IL-4 monoclonal antibody (Product # M3IL4I) at a concentration of 0.005, 0.020, 0.078, 0.312, 1.25, 5 and 20 µg/mL for 90 minutes at 37C. The plate was washed, then incubated with 100 µL per well of an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 62-6520) at a dilution of 1:2000 for 90 minutes at 37C. Detection was performed using 1-Step Ultra TMB substrate (Product # 34028) for 30 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.