- M809 - Invitrogen Antibodies CCL13 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of human Monocyte Chemotactic Protein-4 (MCP-4)/CCL13 was performed by loading 2.5 µg of recombinant human MCP-4 protein per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with StartingBlock (TBS) Blocking Buffer (Product # 37542) for at least 1 hour. The membrane was probed with an MCP4 monoclonal antibody (Product # M809) at a concentration of 2 µg/mL overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:5000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Sandwich ELISA analysis of an anti-human MCP-4 monoclonal antibody (Product # M809) was performed by loading 100 µL per well of antibody (Product # M809) at a concentration of 2 µg/mL overnight at room temperature. The plate was washed 3 times with ELISA Wash Buffer (Product # N503), and 100 µL of recombinant human MCP-4/CCL13 was added to wells in duplicate at 2000, 500, 125, 31.25, 7.813, and 0pg/mL concentrations and the samples were incubated for 2 hours at room temperature. The plate was washed, then incubated with 100 µL per well of a biotinylated MCP-4 monoclonal antibody (Product # M810, biotinylated using EZ-Link Sulfo-NHS-LC-Biotinylation Kit (Product # 21435) at a concentration of 0.13 µg/mL for 1 hour at room temperature, followed by 100 µL per well of Streptavidin-HRP (Product # N504) at a dilution of 1:16,000 for 30 minutes at room temperature. Detection was performed by adding 100 µL of Ultra TMB substrate (Product # 34028) per well and incubating for 30 minutes at room temperature in the dark. The plate was then stopped with 100 µL per well of 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Sandwich ELISA analysis of an anti-human MCP-4 monoclonal antibody (Product # M810) was performed by loading 100 µL per well of antibody (Product # M810) at a concentration of 2 µg/mL overnight at room temperature. The plate was washed 3 times with ELISA Wash Buffer (Product # N503), and 100 µL of recombinant human MCP-4/CCL13 was added to wells in duplicate at 2000, 1000, 500, 250, 125, 62.5, 31.25, and 0pg/mL concentrations and the samples were incubated for 2 hours at room temperature. The plate was washed, then incubated with 100 µL per well of a biotinylated MCP-4 monoclonal antibody (Product # M809, biotinylated using EZ-Link Sulfo-NHS-LC-Biotinylation Kit (Product # 21435) at a concentration of 0.13 µg/mL for 1 hour at room temperature, followed by 100 µL per well of Streptavidin-HRP (Product # N504) at a dilution of 1:16,000 for 30 minutes at room temperature. Detection was performed by adding 100 µL of Ultra TMB substrate (Product # 34028) per well and incubating for 30 minutes at room temperature in the dark. The plate was then stopped with 100 µL per well of 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Sandwich ELISA analysis of an anti-human MCP-4 monoclonal antibody (Product # M810) was performed by loading 100 µL per well of antibody (Product # M810) at a concentration of 2 µg/mL overnight at room temperature. The plate was washed 3 times with ELISA Wash Buffer (Product # N503), and 100 µL of recombinant human MCP-4/CCL13 was added to wells in duplicate at 2000, 1000, 500, 250, 125, 62.5, 31.25, and 0pg/mL concentrations and the samples were incubated for 2 hours at room temperature. The plate was washed, then incubated with 100 µL per well of a biotinylated MCP-4 monoclonal antibody (Product # M809, biotinylated using EZ-Link Sulfo-NHS-LC-Biotinylation Kit (Product # 21435) at a concentration of 0.13 µg/mL for 1 hour at room temperature, followed by 100 µL per well of Streptavidin-HRP (Product # N504) at a dilution of 1:16,000 for 30 minutes at room temperature. Detection was performed by adding 100 µL of Ultra TMB substrate (Product # 34028) per well and incubating for 30 minutes at room temperature in the dark. The plate was then stopped with 100 µL per well of 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.

M809