PA3-045

antibody from Invitrogen Antibodies

Targeting: GPR101

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA3-045

Provider

Invitrogen Antibodies

Product name

GPR101 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Other

Description

Applications Tested: This HTA125 antibody has been pre-diluted and tested by flow cytometric analysis of human TLR4/MD2-transfected cells. This may be used at 5 µL (0.5 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

Conc. Not Determined

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of GPR101 was performed by loading equal amounts of wheat germ lectin agarose bead enriched GPR receptor fractions from mock-transfected or GPR101 transfected HEK293 lysates onto a 7.5% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane, blocked and probed with a GPR101 polyclonal antibody (Product # PA3-045) at a dilution of 1:5000, overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody. Denatured GPR101 was detected at ~64kDa. Chemiluminescent detection was performed using ECL.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of GPR101 was performed on enteric ganglion tissue. To expose target proteins, antigen retrieval was performed by microwaving tissues for 20 minutes in 10mM sodium citrate buffer (pH 6.0). Tissue slides were probed with a GPR101 polyclonal antibody (Product # PA3-045) at a dilution of 1:3000, overnight at 4C in a humidified chamber. Tissues were washed, and detection was performed using an ABC kit composed of biotinylated goat anti-rabbit IgG, peroxidase-conjugated avidin, and 3-amino-9-ethylcarbazole (AEC) substrate in acetate buffer. Tissues were counterstained with hematoxylin and dehydrated to prep for mounting.