PRODUCT CODE: EM1901-62

LAMP2 Monoclonal Antibody (EM1901-62)

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

Western blot analysis of LAMP2 on SiHa cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-62, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of LAMP2 on SiHa cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-62, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
  • ICC staining of LAMP2 in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (EM1901-62, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of LAMP2 in HT-29 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (EM1901-62, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-LAMP2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-62, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human prostate tissue using anti-LAMP2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-62, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-LAMP2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-62, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of LAMP2 was done on SiHa cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1901-62, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of LAMP2 on SiHa cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM1901-62, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

LAMP2 Monoclonal Antibody (EM1901-62)

Immunogen

Recombinant protein within human lamp2 aa 1-300.

Host

Mouse

Positive Control

SiHa cell lysates, A549 cell, HT-29 cell, human liver carcinoma tissue, human prostate tissue, human placenta tissue, SiHa.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

D3-D8-D3

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

2 ug/ul

PURIFICATION

Protein G affinity purified.

MOLECULAR WEIGHT

Predicted band size 45 kDa

Isotype

IgG1

APPLICATION DILUTION

  • WB

  • 1:500-1:2,000

  • ICC

  • 1:50-1:100

  • IHC-P

  • 1:50-1:200

  • FC:1

  • 50-1:100

TARGET

UNIPROT #

PROTEIN NAME

Lysosome-associated membrane glycoprotein 2

GENE NAME

LAMP2

SYNONYMS

LAMP2

SEQUENCE SIMILARITIES

Belongs to the LAMP family.

TISSUE SPECIFICITY

Isoform LAMP-2A is highly expressed in placenta, lung and liver, less in kidney and pancreas, low in brain and skeletal muscle. Isoform LAMP-2B is detected in spleen, thymus, prostate, testis, small intestine, colon, skeletal muscle, brain, placenta, lung, kidney, ovary and pancreas and liver. Isoform LAMP-2C is detected in small intestine, colon, heart, brain, skeletal muscle, and at lower levels in kidney and placenta.

POST-TRANSLATIONAL MODIFICATION

O- and N-glycosylated; some of the 16 N-linked glycans are polylactosaminoglycans.

SUBCELLULAR LOCATION

Cell membrane; Single-pass type I membrane protein. Endosome membrane; Single-pass type I membrane protein. Lysosome membrane; Single-pass type I membrane protein. Cytoplasmic vesicle, autophagosome membrane. Note=This protein shuttles between lysosomes, endosomes, and the plasma membrane.

FUNCTION

Plays an important role in chaperone-mediated autophagy, a process that mediates lysosomal degradation of proteins in response to various stresses and as part of the normal turnover of proteins with a long biological half-live. Functions by binding target proteins, such as GAPDH and MLLT11, and targeting them for lysosomal degradation. Plays a role in lysosomal protein degradation in response to starvation (By similarity). Required for the fusion of autophagosomes with lysosomes during autophagy. Cells that lack LAMP2 express normal levels of VAMP8, but fail to accumulate STX17 on autophagosomes, which is the most likely explanation for the lack of fusion between autophagosomes and lysosomes. Required for normal degradation of the contents of autophagosomes. Required for efficient MHCII-mediated presentation of exogenous antigens via its function in lysosomal protein degradation; antigenic peptides generated by proteases in the endosomal/lysosomal compartment are captured by nascent MHCII subunits. Is not required for efficient MHCII-mediated presentation of endogenous antigens.; [Isoform LAMP-2C]: Modulates chaperone-mediated autophagy. Decreases presentation of endogenous antigens by MHCII. Does not play a role in the presentation of exogenous and membrane-derived antigens by MHCII.