Home \ Antibodies \ GEF-H1, Human, mAb B4/7
Catalog # HM2152

GEF-H1, Human, mAb B4/7

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The monoclonal antibody B4/7 recognizes human and canine guanine nucleotide exchange factor H1 (GEF-H1). GEFH1 is an ~110 kDa protein belonging to the Dbl family of proto-oncogenes. GEF-H1 can activate the small GTPase RhoA, but not Rac1 or Cdc42. Rho family GTPases are central regulators of epithelial tight junctions and the cytoskeleton. GEF-H1 can associate with different cytoskeletal structures, namely microtubules and the actin cytoskeleton. It has also been proposed to mediate crosstalk between the two types of filaments.
Localization of GEF-H1 differs between cell types. In MRC-5 fibroblast cells, GEF-H1 localizes to stress fibers. In epithelial cells, GEF-H1 is associated with apical tight junctions and involved in regulating paracellular permeability of small hydrophilic tracers. Furthermore, its subcellular localization changes in mitotic cells, where endogenous GEF-H1 is concentrated at mitotic spindles. GEF-H1 is capable of binding to the F-actin binding junctional adaptor cingulin. Binding to cingulin inhibits GEF-H1 and results in the downregulation of RhoA and inhibition of G1/S phase transition. In low confluent cultured cells, the localization of GEF-H1 is predominantly cytoplasmic. With increasing density of the cells, free GEF-H1 is sequestered at the tight junctions by cingulin.
GEF-H1 is part of the signaling pathway connecting epithelial polarity with the cell cycle, and as such involved in oncogenesis.

Application
Frozen sections, Immuno fluorescence, Immuno precipitation, Western blot

Application Notes
W: A non-reduced and/or reduced sample treatment and 6-15 % gradient SDS-PAGE was used. The band size is 110 kDa (Ref1-3).
F: Cells were either fixed with methanol at -20  °C or with 3% PFA followed by permeabilization with Triton X-100.

Use
For immunohistochemistry and Western blotting, dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50.

Positive Control
MDCK cells

Product type
Monoclonal antibodies
Amount
100 µg, 20 µg
Formulation
0.2 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin and 0.02% sodium azide.
Immunogen
Protein fraction isolated from detergent extracts of MDCK cells1.
Isotype
Mouse IgG1
Species
Human
Cross reactivity
Canine – Yes
Alias
Rho guanine nucleotide exchange factor 2 (ARHGEF2), Microtubule-regulated Rho-GEF, Proliferating cell nucleolar antigen p40; KIAA0651; LFP40
Storage and stability
Product should be stored at 4 °C. Under recommended storage conditions, product is stable for at least one year. The exact expiry date is indicated on the label.
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result from the use or derivation of this product.
Disease
Neurological disorders, Tumor immunology
Protocol
Protocol Western Blot
59 kb
Protocol SDS Page
57 kb
Protocol Immuno precipitation
54 kb
Protocol Immuno Fluorescence
39 kb
Protocol IHC-F
36 kb
CoA-TDS HM2100-HM2199
HM2152-100UG
79 kb
HM2152-20UG
79 kb
Safety Data Sheet
SDS - HM2152-100UG
52 kb
SDS - HM2152-20UG
52 kb

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