categories: Adaptor Proteins, Adaptor Proteins (Human), Adaptor Proteins (Rodent)

Mouse Monoclonal to NTAL / LAB

NAP-07 (IgG1)

Product No.	Form	Quantity	Price
11-374-C025	purified	0.025 mg	choose region
11-374-C100	purified	0.1 mg	choose region
A4-374-C025	Alexa Fluor® 488	0.025 mg	choose region
A4-374-C100	Alexa Fluor® 488	0.1 mg	choose region
1P-374-C025	PE	0.025 mg	choose region
1P-374-C100	PE	0.1 mg	choose region

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Background

NTAL (Non-T cell activation linker), also known as LAB (Linker for activation of B cells), is a 30 kDa double-palmitoylated transmembrane adaptor protein expressed by B cells, NK cells, mast cells and macrophages. It is a negative regulator of early stages of BCR-dependent B cell signaling and serves as a negative regulator also in mast cells. However, in mast cells, NTAL also contributes to some activation processes, partially overlapping with LAT function.

Entrez Gene (human): 7q11.23
Entrez Gene (mouse): 5 G2

Specificity

The antibody NAP-07 reacts with Non-T cell activation linker (NTAL), also known as LAB (linker of activated B cells), a 25 - 30 kDa transmembrane adaptor protein present in membrane microdomains (rafts) of B lymphocytes, NK cells and myeloid cells.

Immunogen

Recombinant cytoplasmic domain (aa 91-243) of human NTAL.

Species Reactivity: Human Mouse Other not tested Negative Species:	Applications: Flow Cytometry Positive tissue: RAJI human lymphoma cell line RAMOS human lymphoma cell line Immunoprecipitation Western Blotting Recommended dilution: 2 μg/ml, incubation 60 min Positive control: RAMOS human lymphoma cell line Negative control: HeLa human cervix carcinoma cell line Sample preparation: Resuspend approx. 50 mil. cells in 1 ml cold Lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF including Protease inhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate with non-reducing SDS-PAGE sample buffer. Application note: Non-Reducing conditions. 12% separating SDS-PAGE gel. Immunocytochemistry Usage note: Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.	General references Yamasaki S, Ishikawa E, Sakuma M, Kanagawa O, Cheng AM, Malissen B, Saito T.: LAT and NTAL mediate immunoglobulin E-induced sustained extracellular signal-regulated kinase activation critical for mast cell survival. Mol Cell Biol. 2007 Jun;27(12):4406-15. [Abstract] [Full Text] Product Specific References Brdicka T, Imrich M, Angelisova P, Brdickova N, Horvath O, Spicka J, Hilgert I, Luskova P, Draber P, Novak P, Engels N, Wienands J, Simeoni L, Osterreicher J, Aguado E, Malissen M, Schraven B, Horejsi V.: Non-T cell activation linker (NTAL): a transmembrane adaptor protein involved in immunoreceptor signaling. J Exp Med. 2002 Dec 16; 196(12):1617-26. [Abstract] [Full Text] Tkaczyk C, Horejsi V, Iwaki S, Draber P, Samelson LE, Satterthwaite AB, Nahm DH, Metcalfe DD, Gilfillan AM.: NTAL phosphorylation is a pivotal link between the signaling cascades leading to human mast cell degranulation following Kit activation and Fc epsilon RI aggregation. Blood. 2004 Jul 1;104(1):207-14. [Abstract] [Full Text] Volna P, Lebduska P, Draberova L, Simova S, Heneberg P, Boubelik M, Bugajev V, Malissen B, Wilson BS, Horejsi V, Malissen M, Draber P.: Negative regulation of mast cell signaling and function by the adaptor LAB/NTAL. J Exp Med. 2004 Oct 18;200(8):1001-13. [Abstract] [Full Text] *Lebduska P, Korb J, Tůmová M, Heneberg P, Dráber P.: Topography of signaling molecules as detected by electron microscopy on plasma membrane sheets isolated from non-adherent mast cells. J Immunol Methods. 2007 Dec 1;328(1-2):139-51. [Abstract]

Species Reactivity:

Human
Mouse
Other not tested

Negative Species:

Applications:

Flow Cytometry
Positive tissue:
RAJI human lymphoma cell line
RAMOS human lymphoma cell line
Immunoprecipitation
Western Blotting
Recommended dilution: 2 μg/ml, incubation 60 min
Positive control:
RAMOS human lymphoma cell line
Negative control:
HeLa human cervix carcinoma cell line
Sample preparation: Resuspend approx. 50 mil. cells in 1 ml cold Lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF including Protease inhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate with non-reducing SDS-PAGE sample buffer.
Application note: Non-Reducing conditions. 12% separating SDS-PAGE gel.
Immunocytochemistry

Usage note:
Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.

General references

*Yamasaki S, Ishikawa E, Sakuma M, Kanagawa O, Cheng AM, Malissen B, Saito T.: LAT and NTAL mediate immunoglobulin E-induced sustained extracellular signal-regulated kinase activation critical for mast cell survival. Mol Cell Biol. 2007 Jun;27(12):4406-15. [Abstract] [Full Text]

Product Specific References

*Brdicka T, Imrich M, Angelisova P, Brdickova N, Horvath O, Spicka J, Hilgert I, Luskova P, Draber P, Novak P, Engels N, Wienands J, Simeoni L, Osterreicher J, Aguado E, Malissen M, Schraven B, Horejsi V.: Non-T cell activation linker (NTAL): a transmembrane adaptor protein involved in immunoreceptor signaling. J Exp Med. 2002 Dec 16; 196(12):1617-26. [Abstract] [Full Text]

*Tkaczyk C, Horejsi V, Iwaki S, Draber P, Samelson LE, Satterthwaite AB, Nahm DH, Metcalfe DD, Gilfillan AM.: NTAL phosphorylation is a pivotal link between the signaling cascades leading to human mast cell degranulation following Kit activation and Fc epsilon RI aggregation. Blood. 2004 Jul 1;104(1):207-14. [Abstract] [Full Text]

*Volna P, Lebduska P, Draberova L, Simova S, Heneberg P, Boubelik M, Bugajev V, Malissen B, Wilson BS, Horejsi V, Malissen M, Draber P.: Negative regulation of mast cell signaling and function by the adaptor LAB/NTAL. J Exp Med. 2004 Oct 18;200(8):1001-13. [Abstract] [Full Text]

*Lebduska P, Korb J, Tůmová M, Heneberg P, Dráber P.: Topography of signaling molecules as detected by electron microscopy on plasma membrane sheets isolated from non-adherent mast cells. J Immunol Methods. 2007 Dec 1;328(1-2):139-51. [Abstract]

For research use only. Not for drug, diagnostic or other use.

Related Products

Mouse IgG1 Isotype Control

Rabbit Polyclonal to NTAL / LAB

Example Data

NTAL-membrane localization by confocal microscopy

Fig. 1. Confocal microscopy

Fig. 1. Subcellular localization of NTAL by confocal microscopy in THP-1 human acute monocytic leukemia cell line.
THP-1 cells were permeabilized and immunostained using anti-NTAL (NAP-07; green). Nuclei are vizualized by propidium iodide (red).

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