Human LAMP-2/CD107b Antibody

Catalog #: AF6228
2 Citations Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF6228
AF6228-SP

Detection of Human LAMP2/CD107b by Western Blot.

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Citations (2)

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human LAMP-2/CD107b Antibody Summary

Species Reactivity

Human

Specificity

Detects human LAMP2/CD107b in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human LAMP1 is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

Mouse myeloma cell line NS0-derived recombinant human LAMP2/CD107b
Leu29-Phe375
Accession # P13473

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

CyTOF-ready

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

Immunocytochemistry

5-15 µg/mL

See below

Intracellular Staining by Flow Cytometry

2.5 µg/10⁶ cells

See below

Knockout Validated

LAMP‑2/CD107b is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in LAMP‑2/CD107b knockout HeLa cell line.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human LAMP2/CD107b by Western Blot. Western blot shows lysates of JAR human choriocarcinoma cell line, JEG-3 human epithelial choriocarcinoma cell line, and human placenta tissue. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human LAMP2/CD107b Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6228) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). Specific bands were detected for LAMP2/CD107b at approximately 100-120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

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LAMP2/CD107b in HeLa Human Cell Line. LAMP2/CD107b was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human LAMP2/CD107b Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6228) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to lysosomes. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Intracellular Staining by Flow Cytometry

Detection of LAMP-2A antibody in HeLa Human Cell Line antibody by Flow Cytometry.

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Detection of LAMP-2A in HeLa Human Cell Line by Flow Cytometry. HeLa human cervical epithelial carcinoma cell line was stained with Goat Anti-Human LAMP2/CD107b Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6228, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Knockout Validated

Western Blot Shows Human LAMP-2/CD107b Antibody Specificity by Using Knockout Cell Line.

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Western Blot Shows Human LAMP‑2/CD107b Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and LAMP-2/CD107b knockout HeLa cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human LAMP-2/CD107b Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6228) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for LAMP-2/CD107b at approximately 100 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

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: LAMP-2/CD107b

Lysosomal associated membrane protein 2 (LAMP2), also known as CD107b and LGP110, is an approximately 110 kDa transmembrane glycoprotein that is a major component of lysosomal membranes (1). Mature human LAMP2 consists of a 347 amino acid (aa) intralumenal domain, a 24 aa transmembrane segment, and a 35 aa cytoplasmic tail (2). Its lumenal domain is organized into two heavily N-glycosylated regions separated by a Ser/Pro-rich linker that carries a minor amount of O‑linked glycosylation (2, 3). Alternate splicing generates a human LAMP2 isoform (LAMP2B) with a substituted juxtamembrane lumenal region, transmembrane segment, and cytoplasmic tail (4). Within the lumenal domain, human LAMP2 shares approximately 64% aa sequence identity with mouse and rat LAMP2. LAMP2 itself is subject to lysosomal degradation following cleavage of its lumenal domain (5). It mediates the lysosomal uptake of the chaperone HSC73 in complex with cargo proteins and is required for the lysosomal destruction of autophagic vacuoles (6, 7). In cytotoxic T cells and mast cells, LAMP2 is expressed in the membranes of intracellular granules that contain effector molecules such as perforin, granzymes, eicosanoids, and histamine (8-10). Up‑regulated LAMP2 at the plasma membrane serves as an indicator of cell activation of CD8⁺ T cells, mast cells, monocytes, and platelets (9-12). LAMP2 is a native ligand for lectins Galectin-1 and Galectin-3 (13‑15).

References

Eskelinen, E.-L. et al. (2003) Trends Cell Biol. 13:137.
Fukuda, M. et al. (1988) J. Biol. Chem. 263:18920.
Carlsson, S.R. et al. (1988) J. Biol. Chem. 263:18911.
Konecki, D.S. et al. (1995) Biochem. Biophys. Res. Commun. 215:757.
Cuervo, A.M. and J.F. Dice (2000) Traffic 1:570.
Cuervo, A.M. and J.F. Dice (1996) Science 273:501.
Tanaka, Y. et al. (1990) Nature 406:902.
Peters, P.J. et al. (1991) J. Exp. Med. 173:1099.
Betts, M.R. et al. (2003) J. Immunol. Meth. 281:65.
Grutzkau, A. et al. (2004) Cytometry 61:62.
Kannan, K. et al. (1996) Cell. Immunol. 171:10.
Silverstein, R.L. and M. Febbraio (1992) Blood 80:1470.
Skrincosky, D.M. et al. (1993) Cancer Res. 53:2667.
Inohara, H. and Raz, A. (1994) Biochem. Biophys. Res. Commun. 201:1366.
Ohannesian D.W. et al. (1994) Cancer Res. 54:5992.

Long Name

Lysosome-associated Membrane Glycoprotein 2

Entrez Gene IDs

3920 (Human); 16784 (Mouse)

Alternate Names

CD107 antigen-like family member B; CD107b antigen; CD107b; LAMP2; LAMP-2; LAMP2A; LAMP-2A; LAMPB; LGP110; LGP-96; Lysosomal Associated Membrane Protein 2 ; lysosomal-associated membrane protein 2; lysosome-associated membrane glycoprotein 2; Lysosome-associated membrane protein 2

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Citations for Human LAMP-2/CD107b Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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