Mouse splenocytes treated with 10 µg/mL Anti-CD3, 5 µg/mL Anti-CD28, 10 µg/mL Recombinant Human TGF-beta 1 (Catalog # 240-B), and 20 µg/mL Recombinant Mouse IL-2 (Catalog # 402-IL) for 24 hours to induce T regulatory cells (iTregs) were stained with Rat Anti-Mouse CD4 PE-conjugated Monoclonal Antibody (Catalog # FAB554P) and either (A) Rat Anti-Mouse LAP (TGF-beta 1) Monoclonal Antibody (Catalog # MAB7666) or (B) Rat IgG_2AIsotype Control (Catalog # MAB006) followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113).

Applications for Mouse LAP (TGF-beta 1) Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/10⁶ cells
Sample: Mouse splenocytes treated with Anti-CD3, Anti-CD28, Recombinant Human TGF‑ beta1 (Catalog # 240-B), and Recombinant Mouse IL‑2 (Catalog # 402-IL) for 24 hours to induce T regulatory cells (iTregs)

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 4 using MAB7666 in the following applications:

ELISA (1 Review)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. 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.

Background: LAP (TGF-beta 1)

TGF-beta 1 (Transforming Growth Factor beta 1) is one of three closely related mammalian members of the large TGF-beta superfamily that share a characteristic cystine knot structure (1‑7). TGF-beta 1, -2 and -3 are highly pleiotropic cytokines that are proposed to act as cellular switches that regulate processes such as immune function, proliferation and epithelial-mesenchymal transition (1‑4). Each TGF-beta isoform has some non‑redundant functions; for TGF-beta 1, mice with targeted deletion show defects in hematopoiesis and endothelial differentiation, and die of overwhelming inflammation (2). Human TGF‑ beta1 cDNA encodes a 390 amino acid (aa) precursor that contains a 29 aa signal peptide and a 361 aa proprotein (8). A furin‑like convertase processes the proprotein to generate an N‑terminal 249 aa Latency‑Associated Peptide (LAP) and a C‑terminal 112 aa mature TGF‑ beta1 (8, 9). Disulfide‑linked homodimers of LAP and TGF‑ beta1 remain non‑covalently associated after secretion, forming the small latent TGF‑ beta1 complex (8‑10). Covalent linkage of LAP to one of three latent TGF‑ beta binding proteins (LTBPs) creates a large latent complex that may interact with the extracellular matrix (9, 10). TGF‑ beta is activated from latency by pathways that include actions of the protease plasmin, matrix metalloproteases, thrombospondin 1 and a subset of integrins (10). Mature human TGF‑ beta1 shares 100% aa identity with pig, dog and cow TGF‑ beta1, and 99% aa identity with mouse, rat and horse TGF‑ beta1. It demonstrates cross‑species activity (1). TGF‑ beta1 signaling begins with high‑affinity binding to a type II ser/thr kinase receptor termed TGF‑ beta RII. This receptor then phosphorylates and activates a second ser/thr kinase receptor, TGF‑ beta RI, also known as Activin Receptor‑Like Kinase 5(ALK‑5), or alternatively, ALK‑1. This complex phosphorylates and activates Smad proteins that regulate transcription (3, 11, 12). Contributions of the accessory receptors Betaglycan (also known as TGF‑ beta RIII) and Endoglin, or use of Smad‑independent signaling pathways, allow for disparate actions observed in response to TGF‑ beta in different contexts (11).

References

Derynck, R. and K. Miyazono (2008) Cold Spring Harbor Laboratory Press, 29.
Dunker, N. and K. Krieglstein (2000) Eur. J. Biochem. 267:6982.
Wahl, S.M. (2006) Immunol. Rev. 213:213.
Chang, H. et al. (2002) Endocr. Rev. 23:787.
Lin, J.S. et al. (2006) Reproduction 132:179.
Hinck, A.P. et al. (1996) Biochemistry 35:8517.
Mittl, P.R.E. et al. (1996) Protein Sci. 5:1261.
Derynck, R. et al. (1985) Nature 316:701.
Miyazono, K. et al. (1988) J. Biol. Chem. 263:6407.
Oklu, R. and R. Hesketh (2000) Biochem. J. 352:601.
de Caestecker, M. et al. (2004) Cytokine Growth Factor Rev. 15:1.
Zuniga, J.E. et al. (2005) J. Mol. Biol. 354:1052.

Long Name

Latency-associated Peptide

Alternate Names

LAP (TGFbeta 1)

Entrez Gene IDs

7040 (Human)

Gene Symbol

TGFB1

UniProt

P04202

Additional LAP (TGF-beta 1) Products

Product Documents for Mouse LAP (TGF-beta 1) Antibody

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COA

SDS

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Product Specific Notices for Mouse LAP (TGF-beta 1) Antibody

For research use only

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Flow Cytometry

Mouse LAP (TGF-beta 1) Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB7666

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Scientific Data Images for Mouse LAP (TGF-beta 1) Antibody

Detection of LAP (TGF‑ beta1) in Mouse iTreg cells by Flow Cytometry.