M550B

antibody from Invitrogen Antibodies

Targeting: IL5 EDF, IL-5, TRF

Western blot (Recommended by provider)

ELISA (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

M550B

Provider

Invitrogen Antibodies

Product name

IL-5 Monoclonal Antibody (39D10), Biotin

Antibody type

Monoclonal

Antigen

Recombinant full-length protein

Description

M550B targets IL-5 in ELISA, and WB applications and shows reactivity with Human samples. The M550B immunogen is recombinant human IL-5. M550B detects IL-5 which has a predicted molecular weight of approximately 13 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. The M550B IL1b antibody (biotinylated conjugate of clone 39D10) has successfully been paired as the detection antibody in a sandwich ELISA with capture antibody M551 (clone 5A10). Typical dilutions for sandwich ELISA include 1 µg/mL for coating and 0.125 - 0.25 µg/mL for detection. Biotinylated antibody M550B (clone 39D10) and antibody M551 (clone 5A10) have successfully been used in combination with recombinant IL5 protein SIL5 in ELISA applications.

Reactivity

Human

Host

Rat

Conjugate

Biotin

Isotype

IgG

Antibody clone number

39D10

Vial size

250 µg

Concentration

0.5 mg/mL

Storage

-20°C

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Sandwich ELISA analysis of human IL-5 was performed by coating wells of a 96-well plate with 110 µL of a human IL-5 monoclonal antibody (Product # M551) in duplicate at 1, 2, 4, 5, 7, and 9 µg/mL for at least 12 hours. The plate was blocked with a proprietary blocking solution for 1-4 hours at room temperature, and 50 µL of mouse IL-5 recombinant protein (Product # SMIL5) was added in replicates of twelve at 400, 160, 64, 25.6, 10.24, and 0pg/mL for 1 hour at room temperature. The plate was washed and incubated 50 µL of a biotinylated IL-5 monoclonal antibody (Product # M550B) was added to all test wells at 0.5 µg/mL for 1 hour at room temperature. The plate was washed, then incubated with 100 µL per well of Streptavidin-HRP (Product # N504) at a 1:10,000 dilution for 30 minutes at room temperature. Detection was performed with 100 µL per well of Ultra TMB substrate (Product # 34028) for 30 minutes at room temperature in the dark. The plate was then stopped with 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.

Conjugate

Biotin