LAMP-2 / CD107b Antibody

LAMP-2 / CD107b Polyclonal Antibody for Western blot, IF, IHC

>> See 19 other antibodies for LAMP2 (CD107b, LAMP-2, LAMPB, LGP110, CD107 antigen-like family member B, lysosome-associated membrane glycoprotein 2, lysosome-associated membrane protein 2)
Synonyms:
CD107b, LAMP-2, LAMPB, LGP110, CD107 antigen-like family member B, lysosome-associated membrane glycoprotein 2, lysosome-associated membrane protein 2
Entrez Gene ID:
Details
Host / Isotype: Rabbit
Class: Polyclonal
Type: Antibody
Tested Species Reactivity: Human (Hu), Mouse (Ms), Rat (Rt)
Published Species Reactivity: Human (Hu), Mouse (Ms), Rat (Rt)
Immunogen: Synthetic Peptide: C G(400) L K R H H T G Y E Q F(411)
Ordering Information
Pierce LAMP-2 / CD107b Antibody
Product #
PA1-655
Size
100 µL
Price
$324.00
Purchase
Tested Applications Dilution *
Western Blot (WB) 1:100 - 1:1000
Immunofluorescence (IF) Assay dependent
Immunohistochemistry (IHC) 1:20
Published Applications Dilution
Western Blot (WB) See publications below
Immunocytochemistry (ICC) See publications below
Immunohistochemistry (IHC) See publications below
* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
Form Information
Form: Liquid
Concentration: 1mg/ml
Purification: Antigen affinity chromatography
Storage Buffer: PBS with 1mg/ml BSA
Preservative: 0.05% sodium azide
Storage Conditions: -20° C, Avoid Freeze/Thaw Cycles
Product Specific Information
PA1-655 detects lysosome-associated membrane protein-2 (LAMP2) from rat, mouse and human tissues.

PA1-655 has been successfully used in Western blot, immunofluorescence, and immunohistochemistry. By Western blot, this antibody detects glycosylated and non-glycosylated LAMP-2, depending on the lysis buffer used.

PA1-655 immunizing peptide corresponds to amino acid residues 400-411 from rat LAMP2. This sequence is completely conserved between rat, mouse, and guinea pig LAMP2. PA1-655 immunizing peptide (Cat. # PEP-039) is available for use in neutralization and control experiments.
General Information
Lysosomes are membrane-bound vacuoles which play a critical role in cellular metabolism. These organelles participate in endocytosis, the synthesis and assembly of acid hydrolases, act as sites for digestion of foreign materials and for specialized autolytic cellular processes. Membrane-associated glycoproteins have an important role in normal lysosomal function. Prior to posttranslational modification, Lysosome-associated membrane protein-2 (LAMP-2) is an ~45 kDa polypeptide. Mature, functional LAMP2 is extensively glycosylated with a variety of different N-linked and O-linked oligosaccharides with a total molecular weight of ~100-110 kDa. LAMP2 has a structure comprised of a large amino-terminal intra-lysosomal domain, a hydrophobic trans-membrane domain, and a short carboxyl-terminal cytoplasmic tail.
Product Images
  • LAMP-2 / CD107b Antibody (PA1-655) in WB
    LAMP2-Antibody-PA1-655-Western_Blot-1_20150120150820.jpg

    Western Blot with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655) (LAMP2-Antibody-PA1-655-Western_Blot-1_20150120150820.jpg)

    Western Blot with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655)

    Western blot analysis of LAMP-2 was performed by loading 25ug of NRK cell lysates prepared with indicated lysis buffers, and 10ul of PageRuler Plus Prestained Protein Ladder (Product # 26619) onto 4-20% Tris-HCl polyacrylamide gels. Proteins were transferred to PVDF membranes using the G2 Fast Blotter (Product # 62288), and blocked with 5% milk in TBST for 1 hour at room temperature. Before probing the blots, the LAMP-2 polyclonal antibody (Product # PA1-655) was either pre-incubated with a synthetic LAMP-2 peptide (Product # PEP-039) for 1 hour at room temperature (left panel) to block the antigen-binding sites, or was left unincubated (right panel). Both glycosylated and non-glycosylated LAMP-2 were detected after probing with the unblocked LAMP-2 polyclonal antibody at a dilution of 1:1000 in blocking buffer (right panel), whereas only a faint nonspecific band was detected after probing with the LAMP-2 polyclonal antibody pre-incubated with the blocking peptide, also at a dilution of 1:1000 in blocking buffer (left panel) overnight at 4C. Both blots were washed in TBST, and probed with an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:20,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076). Note: Triton X-100 lysis buffer contains 25mM HEPES pH7.5, 150mM NaCl, 1% Triton X-100 and 5mM EDTA.
  • LAMP-2 / CD107b Antibody (PA1-655) in WB
    LAMP2-Antibody-PA1-655-Western_Blot-2_20150120150851.jpg

    Western Blot with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655) (LAMP2-Antibody-PA1-655-Western_Blot-2_20150120150851.jpg)

    Western Blot with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655)

    Western blot analysis of LAMP-2 was performed by loading 10ug of HeLa (left panel), NIH/3T3 (middle panel) and NRK (right panel) whole cell lysates prepared using the Subcellular Protein Fractionation kit (Product # 78840) per well, and 10ul of PageRuler Plus Prestained Protein Ladder (Product # 26619) onto 4-20% Tris-HCl polyacrylamide gels. Proteins were transferred to PVDF membranes using the G2 Fast Blotter (Product # 62288), and blocked with 5% milk in TBST for 1 hour at room temperature. Glycosylated LAMP-2 was detected at ~72kDa after probing with a LAMP-2 polyclonal antibody (Product # PA1-655) at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform, washing in TBST, and probing with an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:20,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).
  • LAMP-2 / CD107b Antibody (PA1-655) in IF
    PA1-655_IF_Hela.jpg

    Immunofluorescence with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655) (PA1-655_IF_Hela.jpg)

    Immunofluorescence with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655)

    Immunofluorescent analysis of LAMP2 in HeLa Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a LAMP2 polyclonal antibody (Product # PA1-655) at a dilution of 1:20 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503). LAMP2 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.
  • LAMP-2 / CD107b Antibody (PA1-655) in IF
    PA1-655_IF_HepG2.jpg

    Immunofluorescence with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655) (PA1-655_IF_HepG2.jpg)

    Immunofluorescence with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655)

    Immunofluorescent analysis of LAMP2 in HepG2 Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a LAMP2 polyclonal antibody (Product # PA1-655) at a dilution of 1:100 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503). LAMP2 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.
  • LAMP-2 / CD107b Antibody (PA1-655) in IF
    LAMP2-Antibody-PA1-655-Immunofluorescence_20150120150733.jpg

    Immunofluorescence with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655) (LAMP2-Antibody-PA1-655-Immunofluorescence_20150120150733.jpg)

    Immunofluorescence with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655)

    Immunofluorescent analysis of LAMP-2 (green) in NIH/3T3 cells. Cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 in PBS, and blocked with 1% Blocker BSA (Product # 37525), each for 15 minutes at room temperature. Cells were stained with a LAMP-2 polyclonal antibody (Product # PA1-655) at a concentration of 20ug/ml in 1% Blocker BSA in PBS (right panel), or incubated in blocking buffer alone as a negative control (left panel) overnight at 4C, and then incubated with a Dylight 488-conjugated goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:1000 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI (Product # 46190). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
  • LAMP-2 / CD107b Antibody (PA1-655) in IHC
    PA1-655_Immunohistochemistry_Placenta-tissue.jpg

    Immunohistochemistry with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655) (PA1-655_Immunohistochemistry_Placenta-tissue.jpg)

    Immunohistochemistry with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655)

    Immunohistochemistry was performed on normal deparaffinized Human placenta tissue tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a rabbit polyclonal antibody recognizing LAMP2 (PA1-655) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
  • LAMP-2 / CD107b Antibody (PA1-655) in IHC
    PA1-655_Immunohistochemistry_Prostate-carcinoma.jpg

    Immunohistochemistry with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655) (PA1-655_Immunohistochemistry_Prostate-carcinoma.jpg)

    Immunohistochemistry with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655)

    Immunohistochemistry was performed on cancer biopsies of deparaffinized Human prostate carcinoma tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a rabbit polyclonal antibody recognizing LAMP2 (PA1-655) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
  • LAMP-2 / CD107b Antibody (PA1-655) in IHC
    PA1-655_Immunohistochemistry_Tonsil-tissue.jpg

    Immunohistochemistry with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655) (PA1-655_Immunohistochemistry_Tonsil-tissue.jpg)

    Immunohistochemistry with anti-LAMP-2 / CD107b Polyclonal Antibody (PA1-655)

    Immunohistochemistry was performed on normal deparaffinized Human tonsil tissue tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a rabbit polyclonal antibody recognizing LAMP2 (PA1-655) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
Publications:
Western Blot
  Species / Dilution Summary
  Hu / Not Cited
PA1-655 was used in western blot to investigate the anti-apoptotic mechanism of Hsp70 in human melanocytes

Carcinogenesis. 2007 Mar;28(3):537-44.
"Hsp70 protects against UVB induced apoptosis by preventing release of cathepsins and cytochrome c in human melanocytes."
Author(s): Bivik C, Rosdahl I, Ollinger K
Number of Citations: 1
(See PubMed article )
  Hu / 1:200
PA1-655 was used in western blot to identify and characterize beta hexosaminidases

Biosci Rep. 2008 Aug;28(4):229-37.
"Identification and characterization of mature beta-hexosaminidases associated with human placenta lysosomal membrane."
Author(s): Magini A, Mencarelli S, Tancini B, Ciccarone V, Urbanelli L, Hasilik A, Emiliani C
Number of Citations: 1
(See PubMed article )
  Ms / 0
PA1-655 was used in western blot to study the role of mitochondria and lysosomes in calpain-mediated epithelial-cell death during mammary gland involution

Cell Death Differ. 2012 Sep;19(9):1536-48.
"Calpains mediate epithelial-cell death during mammary gland involution: mitochondria and lysosomal destabilization."
Author(s): Arnandis T, Ferrer-Vicens I, García-Trevijano ER, Miralles VJ, García C, Torres L, Viña JR, Zaragozá R
Number of Citations: 0
(See PubMed article )
  Rt / Not Cited
PA1-655 was used in western blot to identify membrane-associated RING-CH (MARCH) and its binding to syntaxin 6.

Mol Biol Cell. 2005 Apr;16(4):1696-710.
"MARCH-II is a syntaxin-6-binding protein involved in endosomal trafficking."
Author(s): Nakamura N, Fukuda H, Kato A, Hirose S
Number of Citations: 1
(See PubMed article )
  Rt / Not Cited
PA1-655 was used in western blot to investigate the influence of chronic alcohol consumption on neuronal endocytosis

Toxicol Sci. 2010 May;115(1):202-13.
"Endocytosis in cultured neurons is altered by chronic alcohol exposure."
Author(s): Marín MP, Esteban-Pretel G, Ponsoda X, Romero AM, Ballestín R, López C, Megías L, Timoneda J, Molowny A, Canales JJ, Renau-Piqueras J
Number of Citations: 0
(See PubMed article )
Immunocytochemistry
  Species / Dilution Summary
  Hu / Not Cited
PA1-655 was used in immunocytochemistry to investigate the relationship between macroautophagy and apoptosis in mammalian cells.

Mol Cell Biol. 2005 Feb;25(3):1025-40.
"Inhibition of macroautophagy triggers apoptosis."
Author(s): Boya P, González-Polo RA, Casares N, Perfettini JL, Dessen P, Larochette N, Métivier D, Meley D, Souquere S, Yoshimori T, Pierron G, Codogno P, Kroemer G
Number of Citations: 1
(See PubMed article )
  Ms / Not Cited
PA1-655 was used in immunocytochemistry to investigate the trafficking pathway of EtxB-peptide conjugates.

J Biol Chem. 2004 Dec 3;279(49):51315-22.
"Trafficking of exogenous peptides into proteasome-dependent major histocompatibility complex class I pathway following enterotoxin B subunit-mediated delivery."
Author(s): Hearn AR, de Haan L, Pemberton AJ, Hirst TR, Rivett AJ
Number of Citations: 1
(See PubMed article )
Immunohistochemistry
  Species / Dilution Summary
  Ms / Not Cited
PA1-655 was used in immunohistochemistry to study the mechanism for the subcellular distributions of lysosome-associated membrane protein LAMP-2 splice variants

J Cell Biol. 1997 Jun 2;137(5):1161-9.
"Different steady state subcellular distributions of the three splice variants of lysosome-associated membrane protein LAMP-2 are determined largely by the COOH-terminal amino acid residue."
Author(s): Gough NR, Fambrough DM
Number of Citations: 13
(See PubMed article )
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