- Clone
- W18253B (See other available formats)
- Regulatory Status
- RUO
- Other Names
- G Protein-Coupled Receptor Kinase Interacting, ArfGAP 1, GRK-Interacting Protein 1, ARF GAP GIT1, CAT1, Cool-associated and tyrosine phosphorylated protein 1, ARF GTPase-activating protein GIT1
- Isotype
- Rat IgG2a, κ
- Ave. Rating
- Submit a Review
- Product Citations
- publications
![W18253B_PURE_GIT1_Antibody_1_022120 W18253B_PURE_GIT1_Antibody_1_022120](https://d1spbj2x7qk4bg.cloudfront.net/Admin/Public/GetImage.ashx?Image=/Files/Images/media_assets/products/product_images/W18253B_PURE_GIT1_Antibody_1_022120.png&Width=240&Height=300&altFmImage_path=&Compression=90&Crop=5)
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Whole cell extracts (15 µg protein) from the indicated cell lines were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:500 dilution) of purified anti-GIT1 antibody (clone W18253B) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:50000 dilution (lower). Lane M: Molecular weight marker. Cell lysates were loaded in order of ascending GIT1 expression; predicted expression data was obtained from Human Protein Atlas. -
A549 cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with Triton X-100 for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with 5.0 µg/mL (1:100 dilution) of either purified rat IgG2a, κ isotype ctrl antibody (Cat. No. 400502) (panel A) or purified anti-GIT1 antibody (panel B) overnight at 4°C, followed by incubation with Alexa Fluor® 594 goat anti-rat IgG antibody (Cat. No. 405422) at 2.0 µg/mL. Nuclei were counterstained with DAPI, and the image was captured with a 60X objective. -
Whole cell extracts (250 μg total protein) prepared from A549 cells were immunoprecipitated overnight with 2 μg of purified rat IgG2a, κ isotype ctrl antibody (Cat. No. 400502) or purified anti-GIT1 antibody (clone W18253B). The resulting IP fractions and whole cell extract input (6%) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with a 1 µg/mL (1:500 dilution) of W18253B. Lane M: Molecular weight marker.
Cat # | Size | Price | Quantity Check Availability | Save | ||
---|---|---|---|---|---|---|
935601 | 25 µg | $101 | ||||
935602 | 100 µg | $253 |
GIT1 is a G protein-coupled receptor kinase-interacting protein which is a component of the PIX/Rac/PAK pathway that is involved in cellular development. In this pathway, Rac activates PAK which leads to the phosphorylation of PIX and GIT1. The downstream result to these interactions leads to the formation of focal adhesion complexes in the periphery of the cell membrane with various proteins including FAK and paxillin. GIT1's various protein-protein interactions in the focal complex lead to downstream cellular events such as migration, morphogenesis and neuronal growth (spine/synapse formation).
Product DetailsProduct Details
- Verified Reactivity
- Human, Mouse
- Antibody Type
- Monoclonal
- Host Species
- Rat
- Immunogen
- Partial recombinant human GIT1 protein of approximately 250 amino acids surrounding amino acid 500
- Formulation
- Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
- Preparation
- The antibody was purified by affinity chromatography.
- Concentration
- 0.5 mg/mL
- Storage & Handling
- The antibody solution should be stored undiluted between 2°C and 8°C.
- Application
-
WB - Quality tested
ICC, IP - Verified - Recommended Usage
-
Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.5 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. For immunoprecipitation, the suggested use of this reagent is 2.0 µg/test. It is recommended that the reagent be titrated for optimal performance for each application.
- Application Notes
-
- Non-specific bands present in HeLa lysates.
- Triton X-100 is the best ICC permeabilization method for this antibody.
- RRID
-
AB_2832898 (BioLegend Cat. No. 935601)
AB_2832899 (BioLegend Cat. No. 935602)
Antigen Details
- Structure
- GIT1 is a 761 amino acid protein with a predicted molecular weight of 84 kD.
- Distribution
-
Cytosol and focal adhesion sites
- Function
- GTPase activation
- Biology Area
- Cell Biology, Cell Motility/Cytoskeleton/Structure, Protein Trafficking and Clearance
- Molecular Family
- GTPases, Scaffold Proteins
- Antigen References
-
- Zhang H, et al. 2005. J. Neurosci. 25:3379.
- Zhao ZS, et al. 2000. Mol. Cell. Biol. 20:6354.
- Goehler H, et al. 2004. Mol. Cell. 15:853.
- Gene ID
- 28964 View all products for this Gene ID
- UniProt
- View information about GIT1 on UniProt.org
Related Pages & Pathways
Pages
Related FAQs
Other Formats
View All GIT1 Reagents Request Custom ConjugationDescription | Clone | Applications |
---|---|---|
Purified anti-GIT1 | W18253B | WB,ICC,IP |
Compare Data Across All Formats
This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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