PA3-021

antibody from Invitrogen Antibodies

Targeting: GPR32 RVDR1

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA3-021

Provider

Invitrogen Antibodies

Product name

GPR32 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Other

Description

IHC (P) analysis shows positive staining of GPR32 in human small intestine tissue neuroendocrine cells.

Concentration

Conc. Not Determined

References

Submitted references

Passive and active inclusion of host proteins in human immunodeficiency virus type 1 gag particles during budding at the plasma membrane.

Hammarstedt M, Garoff H
Journal of virology 2004 Jun;78(11):5686-97

---

Differential display implicates cyclophilin A in adult cortical plasticity.

Arckens L, Van der Gucht E, Van den Bergh G, Massie A, Leysen I, Vandenbussche E, Eysel UT, Huybrechts R, Vandesande F
The European journal of neuroscience 2003 Jul;18(1):61-75

---

Interaction of the retinoblastoma gene product, RB, with cyclophilin A negatively affects cyclosporin-inhibited NFAT signaling.

Cui Y, Mirkia K, Florence Fu YH, Zhu L, Yokoyama KK, Chiu R
Journal of cellular biochemistry 2002;86(4):630-41

---

The hydrophobic pocket of cyclophilin is the binding site for the human immunodeficiency virus type 1 Gag polyprotein.

Braaten D, Ansari H, Luban J
Journal of virology 1997 Mar;71(3):2107-13

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of GPR32 was performed by loading equal amounts of wheat germ lectin agarose bead enriched GPR receptor fractions from GPR32 or GPR39 transfected HEK293 lysates and 10 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane using the G2 Fast Blotter (Product # 62288), and blocked with StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 1 hour at room temperature. GPR32 was detected at ~60 kDa using a GPR32 polyclonal antibody (Product # PA3-021) at a dilution of 1:1000 in StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:20,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075). Images were acquired on a Thermo Scientific myECL Imager (Product # 62236).

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of GPR32 was performed on neuroendocrine cells in human small intestine 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 GPR32 polyclonal antibody (Product # PA3-021) 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.