Detection of Human Thymosin beta 4 by Western Blot.
Western blot shows lysates of human peripheral blood lymphocytes (PBL), HL‑60 human acute promyelocytic leukemia cell line, and human mature dendritic cells. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human Thymosin beta 4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6796) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Thymosin beta 4 at approximately 5 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Thymosin beta 4 in HeLa Human Cell Line.
Thymosin beta 4 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human Thymosin beta 4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6796) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red, upper panel; Catalog # NL010) and counterstained with DAPI (blue, lower panel). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Thymosin beta 4
Thymosin beta 4 (T beta 4; also TB4X and Fx) is a 5.0 kDa member of the beta -thymosin family of molecules. Members of this family range from 41-44 amino acids (aa) in length, and possess an isoelectric point that lies between pH 4.0-7.0 ( alpha -thymosins have values less than 4.0). Multiple cell types produce T beta 4, either constitutively, or after stimulation. They include platelets, endothelial cells, neutrophils, astrocytes and macrophages. T beta 4 is both a secreted and intracellular molecule. The secreted form contributes to wound healing and angiogenesis, and may act on ATPase. Intracellularly, it forms a 1:1 complex with G-actin and blocks F-actin polymerization. This regulates the availablility of actin monomers for filament formation and subsequent cell migration. Mature human T beta 4 is 43 aa in length (aa 2-44). It contains an actin-binding site (aa 17-23), one acetylated Ser and five acetylated lysines (4; 12; 26; 32; 39) and one phosphorylation site at Thr23. T beta 4 undergoes proteolytic processing to generate an N-terminal acetylated peptide (aa 2-5: SerAspLysPro). Mature human T beta 4 is identical to mouse T beta 14 in aa sequence, and it shares 74% aa identity with its human family member T beta 10.
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