PA3-046

antibody from Invitrogen Antibodies

Targeting: GPR176 Gm1012

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA3-046

Provider

Invitrogen Antibodies

Product name

GPR176 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Other

Description

GPR176 is in the G protein-coupled receptor family and are cell surface receptors involved in responses to hormones, growth factors, and neurotransmitters. Percent identity with other species by BLAST analysis: Human, Gorilla, Gibbon, Monkey, Marmoset, Mouse, Rat, Hamster, Elephant, Bovine, Dog, Bat, Horse, Stickleback, Zebrafish (100%). GPR176 exists in different glycosylated forms.

Reactivity

Human, Mouse

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 GPR176 was performed by loading equal amounts of wheat germ lectin agarose bead enriched GPR receptor fractions from mock-transfected or GPR176 transfected HEK293 lysates onto a 7.5% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane, blocked and probed with a GPR176 polyclonal antibody (Product # PA3-046) at a dilution of 1:5000, overnight at 4°C on a rocking platform, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody. Denatured GPR176 was detected at ~95kDa. Chemiluminescent detection was performed using ECL.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of GPR176 was performed by loading 20 µg of Mouse Heart tissue, SH-SY5Y, and A431 lysates and 5 µl of PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BX10). Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% Milk in TBST for 1 hour at room temperature. GPR176 was detected at 57kDa using a GPR176 rabbit polyclonal antibody (Product # PA3-046) at a dilution of 1:2000 in blocking buffer overnight at 4C on a rocking platform, followed by a Goat anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A16110) at a dilution of 1:5000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078).

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of GPR176 was performed on blood vessels of human kidney 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 GPR176 polyclonal antibody (Product # PA3-046) 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.