Sprouty Homolog 2 (Drosophila) (SPRY2) antibody

Antigen: Sprouty Homolog 2 (Drosophila) (SPRY2)

Synonyms: hSPRY2, MGC23039, wu:fk66h11, wu:fk93e11, zgc:110440, hspry2, xspry2, sprouty2, Xsprouty2, sprouty-2, Xtsprouty2, SPRY2, MGC139477, DKFZp459N0719

Clonality: Polyclonal

Host: Rabbit

Reactivity: Human, Cow (Bovine), Mouse (Murine), Rat (Rattus)

Application: ELISA, Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN107035

Additional Information

Alternative name: Sprouty-2

Immunogen: This whole rabbit serum was prepared by repeated immunizations with HisX6 tagged full-length human Sprouty-2 protein.

Format: Liquid (sterile filtered)

Description: Sprouty was originally identified as an inhibitor of Drosophila epidermal growth factor (EGF) and fibroblast growth factor (FGF) receptor signaling during tracheal development. Four isoforms of mammalian sprouty are known (Spry1-Spry4). The role of the mammalian analogs has not been clearly elucidated although it is believed that human Sprouty-2 (hSpry2) may be an inhibitor of cellular migration and proliferation. Spry2 and Spry4 show considerable sequence homology between human and mouse at the C-terminus of the protein. Significant sequence divergence occurs at the N-terminus. Spry2 appears to be abundant in brain, lung and heart tissue, with lesser amounts found in kidney and skeletal muscle.

Specificity: This product was prepared from monospecific antiserum by immunoaffinity chromatography using GST-hSpry2 protein coupled to agarose beads followed exhaustive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum.

Application Details

Application Notes: Note: This antibody reacts with human Sprouty-2 protein and is useful for determining its presence in ELISA, immunoblotting and immunoprecipitation. Approximately 20 ul of antibody is used to immunoprecipitate hSpry2 protein from 1 mg of a HeLa cell lysate in 1 ml. Incubate the antibody for 2 h with the cell lysate. Immunoprecipitate the complex with Protein G agarose. Reactivity in other immunoassays has not yet been fully determined. Recommended Dilutions : This affinity purified antibody was assayed by immunoblot and was found to be reactive at a 1:2,000 dilution. A 38 kDa band corresponding to human Sprouty-2 is observed. Detection occurs using HRP Goat anti-Rabbit IgG and ECL. The researcher should determine optimal titers for other applications. * 38 kDa 1 2 Figure - Affinity purified anti-hSpry2 is shown to detect by immunoblot a 38 kDa band (lane 1) corresponding to human Sprouty-2 in lysates of SW-13 cells (human small-cell adenocarcinoma cell line derived from the adrenal cortex). Lane 2 shows reactivity after the antibody was neutralized with GST-hSpry2. The higher molecular weight band in lane 1 indicated by the * may represent post-translational modification of hSprty2 (palmitoylated and phosphorylated) as reported in the Impagnatiello reference. Detection occurs using a 1:2,000 dilution of antibody followed by 1:4,000 dilution of HRP Goat-a-Rabbit with visualization via ECL. Film exposure approximately 1. Other detection systems will yield similar results. 2 of 2.

Concentration: 0.25 mg/ml (by UV absorbance at 280 nm)

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Storage: Store vial at -20° C. For extended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Dilute only prior to immediate use. Expiration date is one (1) year from date of restoration.

Restrictions: For Research Use only

Images

Affinity purified anti-hSpry2 is shown to detect by immunoblot a 38 kDa band (lane 1) corresponding to human Sprouty-2 in lysates of SW-13 cells (human small-cell adenocarcinoma cell line derived from the adrenal cortex). Lane 2 shows reactivity after the antibody was neutralized with GST-hSpry2. The higher molecular weight band in lane 1 indicated by the * may represent post-translational modification of hSprty2 (palmitoylated and phosphorylated) as reported in the Impagnatiello reference. Detection occurs using a 1:2,000 dilution of antibody followed by 1:4,000 dilution of HRP Goat-a-Rabbit with visualization via ECL. Film exposure approximately 1

Immunoprecipitation of HA-tagged hSpry2 using affinity purified anti-hSpry2 followed by immunoblot detection using monoclonal anti-HA. Lysates from Hela cells with over-expression of HAhSpry2 (lane 1) and with no expression (lane 2) were probed.