M2IL4I

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

M2IL4I

Provider

Invitrogen Antibodies

Product name

IL-4 Monoclonal Antibody (E10022)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

M2IL4I targets IL-4 in ELISA and Western blot applications and shows reactivity with Human samples. The M2IL4I immunogen is recombinant human IL-4 purified from Pichia pastoris. M2IL4I detects IL-4 which has a predicted molecular weight of approximately 15 kDa. Endotoxin levels are less than or equal to 10 µg/mL.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

E10022

Vial size

500 µg

Concentration

0.5 mg/mL

Storage

4° C

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 # M2IL4I) 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 # M2IL4I) at a concentration of 0.0012, 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 # M2IL4I) at a concentration of 0.0012, 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.