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Key Documents

MABN2643

Sigma-Aldrich

Anti-RAPGEF1 (C3G) Antibody, clone 3F6

Synonym(s):

Rap guanine nucleotide exchange factor 1;CRK SH3-binding GNRP;Guanine nucleotide-releasing factor 2;Protein C3G

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About This Item

UNSPSC Code:
12352203
NACRES:
NA.43

biological source

mouse

Quality Level

antibody form

purified antibody

antibody product type

primary antibodies

clone

3F6, monoclonal

mol wt

calculated mol wt 120.55 kDa
observed mol wt ~75 kDa

purified by

using protein G

species reactivity

monkey, rat, human, mouse

packaging

antibody small pack of 100 μL

technique(s)

immunocytochemistry: suitable
immunoprecipitation (IP): suitable
western blot: suitable

isotype

IgG1κ

epitope sequence

Internal

Protein ID accession no.

UniProt accession no.

storage temp.

2-8°C

target post-translational modification

unmodified

Gene Information

human ... RAPGEF1(2889)

General description

Rap guanine nucleotide exchange factor 1 (UniProt: Q13905; also known as CRK SH3-binding GNRP, Guanine nucleotide-releasing factor 2, Protein C3G) is encoded by the RAPGEF1 (also known as GRF2) gene (Gene ID: 2889) in human. RAPGEF1 is a ubiquitously expressed protein that regulates multiple cellular activities and is shown to be essential for early embryonic development in mammals. It is involved in cell proliferation, transformation, cytoskeletal organization, and apoptosis. It is induced during skeletal muscle cell and neuronal cell differentiation. Higher expression of RAPGEF1 is observed in adult skeletal muscle and placenta and in fetal brain and heart. Its phosphorylation at tyrosine 504 enhances its activity as Rap guanine nucleotide exchange factor. It has functions that depend on its catalytic activity as well as protein interaction domain. It contains four SH3 binding motifs (aa 281-292; 451-462; 538-549; and 606-617). It binds to SH3 domain of CRK and GRB2 and transduces signals from CRK to activate RAS. It plays a role in the establishment of basal endothelial barrier function and has a role in nerve growth factor-induced sustained activation of Rap1 and neurite outgrowth. Four isoforms of RAPGEF1 have been described that are produced by alternative splicing. (Ref.: Begum, Z., et al. (2018). Mol. Biol. Rep. 45(6); 1809-1819; Sasi Kumar, K., et al. (2015). Biochim. Biophys. Acta. 1853(10); 2629-2639).

Specificity

Clone 3F6 is a mouse monoclonal antibody that detects Rap guanine nucleotide exchange factor 1 (RAPGEF1). It targets an epitope from the internal region.

Immunogen

GST-tagged recombinant protein fragment corresponding to the Crk binding region of human RAPGEF1 (aa 268-648).

Application

Quality Control Testing

Evaluated by Western Blotting in HEK293 cells transfected with GFP-Crk-binding region (CBR).

Western Blotting Analysis (WB): A 1:500 dilution of this antibody detected RAPGEF1 in HEK293 cells transfected with GFP- Crk-binding region (CBR), but not in cells without GFP-CBR transfection.

Tested Applications

Immunocytochemistry Analysis: A representative lot detected RapGEF1 in Immunocytochemistry application (Begum, Z., et al. (2018). Mol Biol Rep. 45(6):1809-1819).

Immunoprecipitation Analysis: A representative lot immunoprecipitated RapGEF1 in Immunoprecipitation application (Begum, Z., et al. (2018). Mol Biol Rep. 45(6):1809-1819).

Western Blotting Analysis: A representative lot detected RapGEF1 in Western Blotting application (Begum, Z., et al. (2018). Mol Biol Rep. 45(6):1809-1819; Sriram, D., et al. (2020). Sci Rep. 10(1):18838).

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Physical form

Purified mouse monoclonal antibody IgG1 in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Reconstitution

0.5 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommended storage: +2°C to +8°C.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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