LAMP2 Mouse Monoclonal Antibody [D3-D8-D3]
Catalog# EM1901-62
LAMP2 Mouse Monoclonal Antibody [D3-D8-D3]
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WB
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IHC-P
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IF-Cell
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Human
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unconjugated
Overview
Product Name
LAMP2 Mouse Monoclonal Antibody [D3-D8-D3]
Antibody Type
Mouse Monoclonal Antibody
Immunogen
Recombinant protein within human LAMP2 aa 1-300.
Species Reactivity
Human
Validated Applications
WB, IHC-P, IF-Cell
Molecular Weight
Predicted band size: 45 kDa
Positive Control
HeLa cell lysate, Jurkat cell lysate, HepG2 cell lysate, HUVEC cell lysate, JAR cell lysate, HEK-293 cell lysate, THP-1 cell lysate, HUVEC, human kidney tissue, human liver tissue.
Conjugation
unconjugated
Clone Number
D3-D8-D3
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG1
Purification Method
Protein G affinity purified.
Application Dilution
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WB
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1:1,000
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IHC-P
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1:20,000
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IF-Cell
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1:100
Target
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.
Background References
1. Dubois A. et. al. LAMP2 expression dictates azacytidine response and prognosis in MDS/AML. Leukemia. 2019 Jun;33(6):1501-1513.
2. Zheng H. et. al. Lamp2 inhibits epithelial-mesenchymal transition by suppressing Snail expression in HCC. Oncotarget. 2018 Jul 13;9(54):30240-30252.
Sequence Similarity
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, lysosome membrane, endosome membrane, autophagosome membrane.
UNIPROT
Synonyms
CD107 antigen like family member B antibody
CD107 antigen-like family member B antibody
CD107b antibody
LAMP 2 antibody
LAMP 2C antibody
LAMP-2 antibody
LAMP2 antibody
LAMP2_HUMAN antibody
LAMPB antibody
LGP 96 antibody
ExpandImages
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Western blot analysis of LAMP2 on different lysates with Mouse anti-LAMP2 antibody (EM1901-62) at 1/1,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: Jurkat cell lysate
Lane 3: HepG2 cell lysate
Lane 4: HUVEC cell lysate
Lane 5: JAR cell lysate
Lane 6: HEK-293 cell lysate
Lane 7: THP-1 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 45 kDa
Observed band size: 110 kDa
Exposure time: 1 minute 2 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1901-62) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
Western blot analysis of LAMP2 on different lysates with Mouse anti-LAMP2 antibody (EM1901-62) at 1/2,000 dilution.
Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-LAMP2 KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 45 kDa
Observed band size: 110 kDa
Exposure time: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1901-62) at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of HUVEC cells labeling LAMP2 with Mouse anti-LAMP2 antibody (EM1901-62) at 1/100 dilution.
Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-LAMP2 antibody (EM1901-62) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Mouse anti-LAMP2 antibody (EM1901-62) at 1/20,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-62) at 1/20,000 dilution for 1 hour 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 liver tissue with Mouse anti-LAMP2 antibody (EM1901-62) at 1/20,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1901-62) at 1/20,000 dilution for 1 hour 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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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