Human Thymosin beta 4 Antibody

R&D Systems | Catalog # AF6796

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Immunocytochemistry, Simple Western

Cited:

Immunohistochemistry, Western Blot, ELISA Capture

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
View all Thymosin beta 4 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human Thymosin beta 4
Ser2-Ser44
Accession # P62328

Specificity

Detects human Thymosin beta 4 in direct ELISAs and Western blots. In direct ELISAs, approximately 5% cross-reactivity with recombinant human (rh) Thymosin beta 10 and rhThymosin beta 16 is observed.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human Thymosin beta 4 Antibody

Detection of Human Thymosin beta 4 antibody by Western Blot.

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 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

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.

Detection of Human Thymosin  beta 4 by Simple WesternTM.

Simple Western lane view shows lysates of HL‑60 human acute promyelocytic leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for Thymosin  beta 4 at approximately 8 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human Thymosin  beta 4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6796) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). This experiment was conducted under reducing conditions and using the 2-40 kDa separation system.
Detection of Thymosin beta 4 by Western Blot

Detection of Thymosin beta 4 by Western Blot

DUB inhibition induces cofilin dephosphorylation. (A) Analysis of actin FRET efficiencies on ACT cell line pre-treated with 500 nM Latrunculin B or 5 µM CK-666 for 1 h prior addition of 15 µM of WP1130 or b-AP15 for one additional hour. Data are expressed as the percentage of FRET efficiency between DMSO-treated and compound-treated cells. Graphs are mean ± SD of n = 3 independent experiments. (B) Immunoblot analysis showing the effect of WP1130 and b-AP15 (1 h, 15 µM) on the levels of actin sequestering proteins. Membranes were probed with the indicated antibodies and anti-ERK2 as loading control. (C) Effect of increasing amounts of WP1130 and b-AP15 on the phosphorylation of cofilin at Ser3. Membranes were probed with the indicated antibodies and quantified against total cofilin signal (bar graph). Upper panel, bar graphs show actin FRET efficiencies recorded on ACT cells upon the same treatment (mean ± SD of 3 independent experiments). immunoblot analysis of lysates obtained from cells treated as above. (D) Time-course analysis of cofilin Ser3 phosphorylation following treatment with 15 µM of WP1130 and b-AP15 for the indicated time. Membranes were probed with the indicated antibodies and quantified against total cofilin signal (bar graph). Upper panel, the graph shows the corresponding effects on actin FRET efficiencies. (E) WP1130 and b-AP15, but not 8TQ, HBX41108 and MG132, reduce cofilin Ser3 phosphorylation levels. Immunoblotting were performed on lysates from Jurkat cells treated with 15 µM of the indicated compound for 30 min. (F) Immunoblot analysis of cofilin Ser3 phosphorylation on cells treated with 500 nM Latrunculin B or 5 µM CK-666 prior addition of 15 µM of WP1130 and b-AP15 for 30 min. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35069199), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Thymosin beta 4 by Immunohistochemistry

Detection of Thymosin beta 4 by Immunohistochemistry

AAV2/7 causes systemic upregulation of thymosin beta 4 in vivo. (a) BALB/c mice were injected with AAV-LacZ or AAV-Tmsb4x three weeks prior to 10 mg/kg ADR or vehicle (saline) administration and culled 14 days after ADR/saline injection at which point thymosin beta 4 levels were quantified. (b) Liver expression of Tmsb4x mRNA (unpaired t test). (c) Expression of TB4 peptide in the liver. Arrowheads refer to cells with particularly high expression of TB4. Scale bars = 50 µm. (d) Plasma concentration of TB4 (one-way ANOVA with Tukey post hoc test). (e) Whole kidney expression of Tmsb4x gene (one-way ANOVA with Tukey post hoc test). LacZ/saline, n = 9; LacZ/ADR, n = 6; Tmsb4x/ADR, n = 5. Data are presented as the mean ± SEM; *P ≤ 0.05, **P ≤ 0.01 and ***P ≤ 0.001. TB4, Tmsb4x, thymosin beta 4; ADR, Adriamycin; LacZ, beta -Galactosidase; AAV, adeno-associated virus. Image collected and cropped by CiteAb from the following open publication (https://www.nature.com/articles/s41598-022-16287-z), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Thymosin beta 4 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line

Simple Western

10 µg/mL
Sample: HL‑60 human acute promyelocytic leukemia cell line

Western Blot

1 µg/mL
Sample: Human peripheral blood lymphocytes (PBL), HL‑60 human acute promyelocytic leukemia cell line, and human mature dendritic cells

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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.

Alternate Names

TB4X, THYB4, TMSB4, TMSB4X

Entrez Gene IDs

7114 (Human)

Gene Symbol

TMSB4X

UniProt

P62328 (Human)

Additional Thymosin beta 4 Products

Product Documents for Human Thymosin beta 4 Antibody

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Product Specific Notices for Human Thymosin beta 4 Antibody

For research use only

Related Research Areas

Cancer Biomarkers

Citations for Human Thymosin beta 4 Antibody

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