Human LAMP-2/CD107b Antibody

R&D Systems, part of Bio-Techne | Catalog # AF6228

R&D Systems, part of Bio-Techne
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AF6228
AF6228-SP
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Human

Applications

CyTOF-ready, Immunocytochemistry, Intracellular Staining by Flow Cytometry, Knockout Validated, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

View all LAMP-2/CD107b Antibodies »

Product Specifications

Immunogen

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

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human LAMP-2/CD107b Antibody

Detection of Human LAMP2/CD107b antibody by Western Blot.

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.
LAMP2/CD107b antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

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

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.
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Applications for Human LAMP-2/CD107b 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.

Immunocytochemistry

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

Intracellular Staining by Flow Cytometry

2.5 µg/106 cells
Sample: HeLa human cervical epithelial carcinoma cell line fixed with paraformaldehyde and permeabilized with saponin

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.

Western Blot

1 µg/mL
Sample: JAR human choriocarcinoma cell line, JEG‑3 human epithelial choriocarcinoma cell line, and human placenta tissue
Please Note: Optimal dilutions of this antibody should be experimentally determined.

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.

Reconstitution Buffer Available:
Size / Price
Qty

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: 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

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

Long Name

Lysosome-associated Membrane Glycoprotein 2

Alternate Names

CD107b, LAMP2, LAMPB, LGP110

Entrez Gene IDs

3920 (Human); 16784 (Mouse)

Gene Symbol

LAMP2

UniProt

P13473

Additional LAMP-2/CD107b Products

Product Documents for Human LAMP-2/CD107b Antibody

Product Datasheets

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human LAMP-2/CD107b Antibody

For research use only

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