LAMP2 ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Lysosomal Associated Membrane Protein 2 (LAMP2) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Lysosomal Associated Membrane Protein 2 (LAMP2) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

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 (PubMed:8662539, PubMed:11082038, PubMed:18644871, PubMed:24880125, PubMed:27628032) . Functions by binding target proteins, such as GAPDH and MLLT11, and targeting them for lysosomal degradation (PubMed:8662539, PubMed:11082038, PubMed:18644871, PubMed:24880125) . Plays a role in lysosomal protein degradation in response to starvation (By similarity) . Required for the fusion of autophagosomes with lysosomes during autophagy (PubMed:27628032) . 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 (PubMed:27628032) . Required for normal degradation of the contents of autophagosomes (PubMed:27628032) . 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 (PubMed:20518820) . Is not required for efficient MHCII-mediated presentation of endogenous antigens (PubMed:20518820) .

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Lysosomal Associated Membrane Protein 2 (LAMP2) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Lysosomal Associated Membrane Protein 2 (LAMP2) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Lysosomal Associated Membrane Protein 2 (LAMP2), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Lysosomal Associated Membrane Protein 2 (LAMP2) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery Range (%) Average (%) serum (n=5) 79-94 82 EDTA plasma (n=5) 97-104 101 heparin plasma (n=5) 89-97 92 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 79-94 82 serum (n=5) 79-94 82 EDTA plasma (n=5) 97-104 101 EDTA plasma (n=5) 97-104 101 heparin plasma (n=5) 89-97 92 heparin plasma (n=5) 89-97 92

Component Amount Anti-LAMP2 Microplate 96 Wells (12 x 8 Well Strips) LAMP2 Lyophilized Standard 2 x 4 ng 100X Biotinylated LAMP2 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-LAMP2 Microplate 96 Wells (12 x 8 Well Strips) Anti-LAMP2 Microplate 96 Wells (12 x 8 Well Strips) LAMP2 Lyophilized Standard 2 x 4 ng LAMP2 Lyophilized Standard 2 x 4 ng 100X Biotinylated LAMP2 Detector Antibody 120 uL 100X Biotinylated LAMP2 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

This assay has high sensitivity and excellent specificity for detection of Lysosomal Associated Membrane Protein 2 (LAMP2) . No significant cross-reactivity or interference between Lysosomal Associated Membrane Protein 2 (LAMP2) and analogues was observed