Immunofluorescence analysis of GRIP1 was done on 70% confluent log phase PC-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with GRIP1 Rabbit Polyclonal Antibody (PA1846) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Hamster, Human|
|Published species reactivity||Rat, Non-human primate, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues P(206) L P D S E E E G H D S(217) C of human GRIP1.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 3 publications below|
PA1-846 detects glucocorticoid receptor interacting protein-1 (GRIP1) from hamster and human cells.
PA1-846 has been successfully used in immunocytochemistry, and immunofluorescence procedures. Immunocytochemical staining of GRIP1 in CHO cells with PA1-846 results in cytoplasmic and nuclear staining. This antibody has not been used successfully in Western blot procedures.
PA1-846 immunogen is a synthetic peptide corresponding to residues P(206) L P D S E E E G H D S(217) C of human GRIP1. The PA1-846 immunizing peptide (Cat. # PEP-050) is available for use in neutralization and control experiments.
Steroid and thyroid hormones and retinoic acid regulate a complex array of gene expression activity via intracellular receptor transcription factors belonging to the ligand-dependent nuclear receptor superfamily. Adding to the complexity of function of these transcription factors are associated proteins known as coactivators and corepressors which, as their names suggest, enhance or depress transcription activity of the nuclear receptor with which they associate. Glucocorticoid receptor interacting protein-1 (GRIP1), or Transcription Intermediary Factor 2 (TIF2), is a member of a family of transcriptional coactivator proteins which includes steroid receptor coactivator-1 (SRC-1) and Amplified in breast cancer-1 (AIB-1).
GRIP1 has been shown to interact and stimulate transcriptional activity of the retinoic acid, retinoid X, vitamin D, mineralocorticoid, glucocorticoid, thyroid hormone, androgen, estrogen, progesterone, and peroxisome proliferator activated receptors. GRIP1 also contains several copies of the conserved LXXLL/LLXXL motif which has been demonstrated to be critical to receptor-coactivator interactions.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Regulation of GRIP1 and CBP Coactivator activity by Rho GDI modulates estrogen receptor transcriptional enhancement.
PA1-846 was used in western blot to investigate the regulatory mechanism for GRIP1 and CBP coactivator activity.
|Su LF,Wang Z,Garabedian MJ||The Journal of biological chemistry (277:37037)||2002|
|Non-human primate||Not Cited||
Multiple receptor interaction domains of GRIP1 function in synergy.
PA1-846 was used in western blot to study the mechanism for the effect of nuclear hormone receptors on gene transcription
|Schmidt S,Baniahmad A,Eggert M,Schneider S,Renkawitz R||Nucleic acids research (26:1191)||1998|
GRIP1, a transcriptional coactivator for the AF-2 transactivation domain of steroid, thyroid, retinoid, and vitamin D receptors.
PA1-846 was used in western blot to investigate the important role of GRIP1 as a transcriptional coactivator interacting with the hormone binding domain of nuclear hormone receptors
|Hong H,Kohli K,Garabedian MJ,Stallcup MR||Molecular and cellular biology (17:2735)||1997|