FHL2 antibody

Catalog No. ABIN487469

The Mouse Monoclonal anti-FHL2 antibody has been validated for WB, IHC (p) and IP. It is suitable to detect FHL2 in samples from Human and Mouse.

Quick Overview for FHL2 antibody (ABIN487469)

Target: FHL2 (Four and A Half LIM Domains 2 (FHL2))

Reactivity: Human, Mouse

Host: Mouse

Clonality: Monoclonal

Conjugate: This FHL2 antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP)

Clone: 31-10-34

Product Details anti-FHL2 Antibody

Specificity: This antibody reacts with Mouse FHL2.

Cross-Reactivity (Details): Species reactivity (tested):Human and Mouse.

Characteristics: Synonyms: FHL-2, SLIM 3, DRAL, Four and a half LIM domains protein 2, Skeletal muscle LIM-protein 3,LIM domain protein DRAL

Purification: Protein-A Sepharose Chromatography.

Immunogen: His-FHL2. Remarks: Hybridoma was established by fusion of Mouse myeloma cell SP2/0-Ag4 withBalb/c mouse splenocyte.

Isotype: IgG2a

Application Details

Application Notes: Western blot: 1-5 μg/mLPositive Control: C2C12Immunoprecipitation: 3 μg/200-300 μL of cell extract. Positive Control: C2C12Immunohistochemistry: 1-5 μg/mLHeat treatment is necessary for Paraffin Embedded Sections. Autoclave, 10 minutes in citrate buffer ( pH 6.5) at 110 °C

Protocol: SDS-PAGE and Western Blotting1) Wash the cells 3 times with PBS and suspend with 10 volumes of cold Lysis buffer (50mM Tris-HCl, pH 7. 2, 250 mM NaCl, 0. 1% NP-40, 2 mM EDTA, 10% glycerol) containingappropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, thensonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4°C and transfer the supernatant toanother tube. Measure the protein concentration of the supernatant and add the Lysisbuffer to make an 8 mg/mL solution. 3) Mix the sample with an equal volume of Laemmli's sample buffer. 4) Boil the samples for 2 minutes and centrifuge. Load 10 µL of the sample per lane in a 1mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm2 for 1 hourin a semi-dry transfer system. (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for specific transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH7. 2) for 1 hour at RT or overnight at 4°C. 7) Incubate the membrane with the anti-FHL2 (11-134) monoclonal antibody (1-5 µg/mL)diluted with 1% skimmed milk (in PBS, pH 7. 2) for 1 hour at RT. 8) Wash the membrane with PBS (5 minutes x 6 times). 9) Incubate the membrane with the 1: 10000 POD-conjugated anti-mouse IgG diluted with1% skimmed milk (in PBS, pH 7. 2) for 1 hour at RT. 10) Wash the membrane with PBS (5 minutes x 6 times). 11) Wipe excess buffer from the membrane, then incubate it with appropriatechemiluminescence reagents for 1 minute. Remove extra reagent from the membrane bydabbing with a paper towel, and seal it in plastic wrap. 12) Expose to X-ray film in a dark room for 5 minutes. Develop the film as usual. Theconditions for exposure and development may vary. Positive Controls for Western blotting: C2C12Immunoprecipitation1) Wash the cells 3 times with PBS and suspend with 10 volumes of cold Lysis buffer (50mM Tris-HCl, pH 7. 2, 250 mM NaCl, 0. 1% NP-40, 2 mM EDTA, 10% glycerol) containingappropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, then

Restrictions: For Research Use only

Handling

Concentration: 1.0 mg/mL

Buffer: PBS, pH 7.2 containing 50 % Glycerol without preservatives.

Preservative: Without preservative

Storage: -20 °C

Storage Comment: Store the antibody undiluted at -20 °C.
Shelf life: one year from despatch.

Expiry Date: 12 months

Target Details for FHL2

Target: FHL2 (Four and A Half LIM Domains 2 (FHL2))

Alternative Name: FHL2 / SLIM3

Background: Proteins containing LIM domains (which are double zinc finger motifs implicated in protein binding) are important regulators of cell growth, cell differentiation, and remodeling of the cell cytoskeleton. Human Four-and-a-half LIM-only protein 2 (FHL2), also known as DRAL/Slim3 is a 32 kDa protein expressed predominantly in human heart and to a lesser extent in skeletal muscle, testis, and prostate epithelium. Since FHL2 is abundant in heart tissue, it may play a role in the regulation of myofibrillogenesis of heart via LIM-domain binding to focal adhesions. FHL2 has also been identified as a co-activator of the androgen receptor where it promotes androgen receptor transcriptional activity. Stimulation of the Rho signaling pathway induces translocation of FHL2 to the nucleus and subsequent activation of FHL2- and androgen receptor-dependent genes. FHL2 also acts as a trancriptional repressor in muscle cells and is involved in modulation of beta-catenin-dependent transcription of Wnt-responsive genes.Synonyms: DRAL, FHL-2, Four and a half LIM domains protein 2, LIM domain protein DRAL, SLIM 3, Skeletal muscle LIM-protein 3

Gene ID: 2274

UniProt: Q14192

Pathways: Intracellular Steroid Hormone Receptor Signaling Pathway, Regulation of Lipid Metabolism by PPARalpha