- MA1-164 - Invitrogen Antibodies LAMP1 antibody

Submitted references KIF5A-dependent axonal transport deficiency disrupts autophagic flux in trimethyltin chloride-induced neurotoxicity.

Liu M, Pi H, Xi Y, Wang L, Tian L, Chen M, Xie J, Deng P, Zhang T, Zhou C, Liang Y, Zhang L, He M, Lu Y, Chen C, Yu Z, Zhou Z
Autophagy 2021 Apr;17(4):903-924

Extracellular and Luminal pH Regulation by Vacuolar H+-ATPase Isoform Expression and Targeting to the Plasma Membrane and Endosomes.

Smith GA, Howell GJ, Phillips C, Muench SP, Ponnambalam S, Harrison MA
The Journal of biological chemistry 2016 Apr 15;291(16):8500-15

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of LAMP1 was performed by loading NRK cell lysates prepared with indicated lysis buffers and protein amounts per well, and 10 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane using the G2 Fast Blotter (Product # 62288), and blocked with 5% milk in TBST for 1 hour at room temperature. Glycosylated LAMP1 was detected at ~130 kDa after probing with a LAMP1 monoclonal antibody (Product # MA1-164) at a dilution of 1:250 in blocking buffer overnight at 4C on a rocking platform, washing in TBST, and probing with an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of LAMP1 was performed by loading NRK cell lysates prepared with indicated lysis buffers and protein amounts per well, and 10 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane using the G2 Fast Blotter (Product # 62288), and blocked with 5% milk in TBST for 1 hour at room temperature. Glycosylated LAMP1 was detected at ~130 kDa after probing with a LAMP1 monoclonal antibody (Product # MA1-164) at a dilution of 1:250 in blocking buffer overnight at 4C on a rocking platform, washing in TBST, and probing with an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) at a dilution of 1:20,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of LAMP1 (green) in NRK cells. The 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 LAMP1 monoclonal antibody (Product # MA1-164) at a concentration of 10 µg/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-mouse IgG secondary antibody (Product # 35502) 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of LAMP1 (Lysosome-associated membrane glycoprotein 1) was performed using 80% confluent log phase L6 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with LAMP1 Monoclonal Antibody (LY1C6) (Product # MA1-164) at 10 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2500 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing punctate lysosome-like staining in the cytoplasm for LAMP1. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of LAMP1 (Lysosome-associated membrane glycoprotein 1) was achieved by transfecting L6 cells with LAMP1 specific siRNA (Silencer® select Product # s129495, s129497). Immunofluorescence analysis was performed on untransfected L6 cells (panel a to d), transfected with non-specific scrambled siRNA (panels e to h) and transfected with LAMP1 specific siRNA (panel i to l). Cells were fixed, permeabilized, and labelled with LAMP1 Monoclonal Antibody (LY1C6) (Product # MA1-164, 10 µg) followed by Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2500 dilution) (Green). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (Red) staining. Reduction in the number of cytoplasmic punctate staining upon siRNA mediated knockdown (panel i,l) confirms specificity of the antibody to LAMP1. The Images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 2. Expression of V-ATPase subunit isoforms in PC-3 cells. A, extracted mRNA from PC-3 cells was reverse-transcribed and analyzed on Affymetrix DNA microarrays (see ""Experimental Procedures""). Mean values were determined from the output of two independent analyses. Columns indicating isoforms of a particular subunit have equivalent shading . Subunits represented on the array by only a single gene (A, c, D, e, F, H and Ac45) have the same shading . Output for the ATP6V0A1-4 genes is magnified in inset. B, to detect proteins accessible to the extracellular medium, intact PC-3 cells were labeled with membrane-impermeable biotinylation reagent prior to solubilization and extraction with StrepTactin affinity beads, followed by immunoblot analysis ( Surface ). As a control for reactivity, a detergent-solubilized membrane fraction was subjected to the same analysis ( Membranes ). Non-biotinylated protein gave negligible binding to the StrepTactin resin ( Control ). The Membranes samples contain 20 mug of total protein with the exception of the a 4 blot, which contains 50 mug. Surface sample loading is the same in each blot, adjusted to give equivalent intensity in membranes and surface for Ac45. Chemiluminescence capture for a 4 was 10-fold longer than for the other immunoblots. C, knockdown of a 1 , a 3 , and Ac45 expression by RNAi. PC-3 cell lysates (15 mug of protein) were analyzed by immunoblotting ( IB ) with the corresponding subunit-specific antibody after transfecti

MA1-164

Antibody data