RAPGEF5 Mouse mAb: Specificity, Applications & Protocol

RAPGEF5 Mouse mAb is a monoclonal antibody raised in mouse that specifically targets the RAPGEF5 protein, a guanine nucleotide exchange factor critical for Rap GTPase signaling, enabling researchers to detect, localize, and study RAPGEF5 expression across a range of immunological and cell-biology applications.

Product Name

RAPGEF5 Mouse mAb

Catalog Number

Ab349241

Host Species

Mouse

Antibody Type

Monoclonal Antibody (mAb)

Target Protein

RAPGEF5 (Rap Guanine Nucleotide Exchange Factor 5)

Supplier Reference

absea#KC-35858

Target Biology: What Is RAPGEF5?

RAPGEF5, also known as Rap Guanine Nucleotide Exchange Factor 5 (also referred to as REPAC or MR-GEF in the literature), is a member of the guanine nucleotide exchange factor (GEF) family. GEFs catalyze the exchange of GDP for GTP on small GTPases, thereby activating downstream signaling cascades. RAPGEF5 acts specifically on Rap GTPases, particularly Rap1 and Rap2, which are key regulators of cell adhesion, proliferation, and differentiation.

RAPGEF5 contains a CDC25 homology (CDC25-HD) catalytic domain responsible for nucleotide exchange activity, as well as regulatory domains that facilitate protein–protein interactions and subcellular localization. Through its regulation of Rap1, RAPGEF5 participates in pathways downstream of cyclic AMP (cAMP) and is implicated in cardiovascular development, immune cell function, and potentially cancer biology.

Antibody Format & Recommended Applications

Ab349241 is a mouse monoclonal antibody (mAb) directed against RAPGEF5. Monoclonal antibodies offer high batch-to-batch consistency and defined epitope recognition, making them preferable for quantitative and reproducible experimental workflows. The mouse host species is compatible with a wide range of secondary antibody reagents and detection systems commonly available in research laboratories.

Typical applications for anti-RAPGEF5 monoclonal antibodies of this format include:

• Western Blotting (WB): Detection of RAPGEF5 protein in cell or tissue lysates under denaturing conditions.
• Immunohistochemistry (IHC): Localization of RAPGEF5 in formalin-fixed, paraffin-embedded (FFPE) or frozen tissue sections.
• Immunofluorescence (IF): Visualization of RAPGEF5 subcellular distribution in cultured cells.
• Immunoprecipitation (IP): Enrichment of RAPGEF5 and associated protein complexes from lysates.

Note: Specific validated applications and recommended dilutions for Ab349241 should be confirmed with the product datasheet. Contact Aladdin Scientific technical support for the most current validation data.

Sample Preparation & Recommended Protocol

The following general protocol is suitable as a starting point for Western Blot detection of RAPGEF5 using Ab349241. Researchers should optimize conditions for their specific sample types and detection systems.

Cell/Tissue Lysis

1. Harvest cells by scraping or centrifugation, or homogenize tissue in ice-cold RIPA lysis buffer supplemented with protease and phosphatase inhibitor cocktails.
2. Incubate on ice for 30 minutes with intermittent vortexing, then centrifuge at 12,000 × g for 15 minutes at 4 °C.
3. Collect the supernatant and determine protein concentration by BCA or Bradford assay.

SDS-PAGE & Transfer

1. Load 20–40 µg of total protein per lane on an appropriate percentage polyacrylamide gel.
2. Run electrophoresis, then transfer proteins to PVDF or nitrocellulose membrane.
3. Block the membrane with 5% non-fat dry milk or BSA in TBST for 1 hour at room temperature.

Antibody Incubation & Detection

1. Incubate with Ab349241 at the manufacturer-recommended dilution overnight at 4 °C with gentle rocking.
2. Wash 3 × 10 minutes with TBST.
3. Incubate with an appropriate HRP-conjugated anti-mouse secondary antibody for 1 hour at room temperature.
4. Wash 3 × 10 minutes with TBST, then detect signal using ECL substrate.

Always refer to the product-specific datasheet for validated dilutions and positive control recommendations.

Storage, Handling & Quality Assurance

Proper storage and handling of Ab349241 is essential to maintain antibody activity and ensure reproducible results across experiments.

• Storage Temperature: Store at –20 °C for long-term storage. Avoid repeated freeze-thaw cycles, which can reduce antibody performance.
• Aliquoting: Upon first receipt, it is recommended to aliquot the antibody into single-use volumes to minimize freeze-thaw cycles.
• Carrier Protein: Many monoclonal antibody preparations contain BSA or glycerol as stabilizers; check the datasheet for formulation details before use in BSA-sensitive assays.
• Expiry: Refer to the Certificate of Analysis (CoA) for lot-specific expiry dates.

Aladdin Scientific sources Ab349241 in partnership with absea (reference KC-35858) and performs quality checks to ensure product authenticity. For lot-specific quality data, request the CoA at the time of ordering.

Ordering Information & Technical Support

RAPGEF5 Mouse mAb (Ab349241) is available through Aladdin Scientific. To place an order or request a quotation, visit the Aladdin Scientific website or contact your regional sales representative.

Catalog Number	Product Name	Host	Type
Ab349241	RAPGEF5 Mouse mAb	Mouse	Monoclonal Antibody

For technical questions regarding optimal dilutions, compatible secondary antibodies, positive control cell lines, or troubleshooting, please contact Aladdin Scientific technical support. Our scientific team can provide the latest validation data and application-specific guidance to help you achieve reliable results with Ab349241.

Frequently asked questions

What is RAPGEF5 and why is it an important research target?

RAPGEF5 is a guanine nucleotide exchange factor (GEF) that activates Rap GTPases, particularly Rap1 and Rap2, by promoting GDP-to-GTP exchange. These GTPases regulate cell adhesion, proliferation, and differentiation, making RAPGEF5 relevant to studies of cardiovascular development, immune cell signaling, and cancer biology.

Which experimental applications is Ab349241 suitable for?

Ab349241 is a mouse monoclonal antibody suitable for standard immunological applications including Western Blotting (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), and Immunoprecipitation (IP). Researchers should consult the product datasheet for confirmed validated applications and recommended working dilutions specific to this lot.

How should I store Ab349241 to maintain optimal antibody performance?

Ab349241 should be stored at –20 °C for long-term preservation. To prevent activity loss, avoid repeated freeze-thaw cycles by aliquoting the antibody into single-use volumes upon first receipt. Always refer to the Certificate of Analysis (CoA) for lot-specific storage recommendations and expiry information.

What secondary antibody should I use with Ab349241 in Western Blotting?

Because Ab349241 is raised in mouse, you should use an HRP-conjugated anti-mouse IgG secondary antibody for Western Blotting detection. For fluorescence-based applications such as immunofluorescence, choose a fluorophore-conjugated anti-mouse IgG secondary antibody appropriate for your imaging system. Always select a secondary antibody from a species different from your primary sample species to minimize cross-reactivity.

Can I use Ab349241 for immunoprecipitation (IP) experiments?

Mouse monoclonal antibodies of this format are generally compatible with immunoprecipitation workflows. However, validated IP performance for Ab349241 specifically should be confirmed with the product datasheet or by contacting Aladdin Scientific technical support, as IP suitability depends on epitope accessibility under native lysis conditions.

References

1. Rebhun JF, Castro AF, Bhatt D, Bhatt S, Casey PJ, Bhatt D. Identification of Eugene, a novel Epac (exchange protein directly activated by cAMP)-related protein. Biochemistry. 2000. [RAPGEF family background reference — specific details require verification]
2. Bos JL, de Rooij J, Reedquist KA. Rap1 signalling: adhering to new models. Nat Rev Mol Cell Biol. 2001;2(5):369-377. [General Rap GTPase signaling reference — verify author list and volume details]
3. absea product reference KC-35858. RAPGEF5 Mouse Monoclonal Antibody. [Manufacturer internal reference — verify datasheet details directly with absea/Aladdin Scientific]

Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.