14-9778-80

antibody from Invitrogen Antibodies

Targeting: LAMP5 BAD-LAMP, C20orf103, dJ1119D9.3, UNC-46

Provider product page for 14-9778-80

Western blot

ELISA

Immunoprecipitation

Immunohistochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [5]
Comments [0]
Validations
Immunohistochemistry [1]
Other assay [2]

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Product number: 14-9778-80 - Provider product page
Provider: Invitrogen Antibodies
Product name: LAMP5 (BAD-LAMP) Monoclonal Antibody (34.2), eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: The monoclonal antibody 34.2 recognizes human lysosome-associated membrane protein 5 (LAMP5), a member of the LAMP family. LAMP5 differs from other LAMP family members in that its expression is more restricted and is referred to as brain and dendritic cell-associated LAMP-like molecule (BAD-LAMP). In humans, BAD-LAMP is expressed in post-natal neurons and non-activated plasmacytoid dendritic cells. Expression in neurons is localized to intracellular vesicles distributed in specific microdomains along neurites and may play a role in endocytosis. BAD-LAMP is localized to the endoplasmic reticulum-Golgi intermediate compartment (ERGIC) of plasmacytoid dendritic cells and is lost upon TLR activation. This 34.2 antibody has been tested by immunohistochemistry of formalin-fixed paraffin embedded tissue using high or low pH antigen retrieval and can be used at less than or equal to 1 µg/mL. This 34.2 antibody has been tested by western blot analysis of reduced SK-N-SH cell lysates and can be used at less than or equal to 5 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human
Host: Rat
Isotype: IgG
Antibody clone number: 34.2
Vial size: 25 µg
Concentration: 0.5 mg/mL
Storage: 4° C

LAMP5 protein structure - 14-9778-80 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 1 BAD-LAMP is down-modulated after IRF7 activation. a ( left ) Flow cytometry analysis of CAL-1 cells ( top ) and freshly isolated pDCs ( bottom ) from healthy donors stained for both extracellular (CD123 and BADCA4) and intracellular (TLR9 and BAD-LAMP) pDCs markers. ( right ) Flow cytometry histogram plots of intracellular staining for BAD-LAMP, TLR9 and UNC93B1 in both CAL-1 ( left ) and freshly isolated pDCs ( right ) at steady state ( black line ) and after 24 h of CpG-A stimulation ( dashed line ). Full grey histograms represent isotype controls staining. Data are representative of a minimum of three independent experiments. b CAL-1 ( black line ) and freshly isolated pDCs ( dashed line ) were treated with CpG-A for indicated times. BAD-LAMP mRNA levels were measured by RT-qPCR. Raw data have been normalised to housekeeping gene (GAPDH) and graphics represent fold change +- s.d. compared to non-stimulated cells from a minimum of three independent experiments. c IFNalpha 2 ( red ) and TNF ( blue ) mRNA level from CAL-1 ( left ) and freshly isolated pDCs ( right ) were measured by RT-qPCR. Raw data have been normalised to housekeeping gene (GAPDH) and graphics represent fold change +- s.d. compared to non-stimulated cells from a minimum of three independent experiments. d ( top ) CAL-1 cells were treated with CpG-A for indicated times prior lysis and sodium dodecyl sulphate-polyacrylamide gel electrophoresis treatment. Expression of BAD-LAMP, TBK1, IRF7, STAT1, the p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 10 BAD-LAMP expression in tumour-associated pDCs correlates with low IFN-alpha production. a ( left ) Flow cytometry pDC gating strategy on both PBMC ( top ) and tumour ( bottom ) is shown. pDCs (BDCA2 + ; CD123 + ) were segregated from conventional dendritic cells (Lin - , MHC II high ) after identification of live hematopoietic cells (CD45 + Live Dead - ). ( right ) CD123 + /BDCA4 + pDCs from blood ( black ) or from primary breast tumours ( red ) were further analysed for BAD-LAMP and MHC II expression. Data are representative of three patients. b Freshly isolated pDCs from healthy donors were treated for 16 h with tumour supernatant either devoid of (snTUM-) or enriched in (snTUM+) TNF and TGF-beta. ( left ) BAD-LAMP and MHC II histogram plots from FACS staining at steady state ( black line ) or 24 h ( dashed line ) after CpG-A treatment. Full grey histogram represents isotype control. Data are representative for five independent experiments performed with eight different tumour supernatants. ( right ) Levels of BAD-LAMP shown as MFI from FACS intracellular staining in pDCs pre-treated with snTUM- ( dashed line ), snTUM + ( red line ) or with medium ( black lines ). MFI +- s.d. from five independent experiments. c Freshly isolated pDCs from heathy donors were treated for 16 h with tumour supernatant either devoid of (snTUM-; black) or enriched in (snTUM + ; red) in TNF and TGF-beta. 2D FACS analysis of intracellular staining for IFN-alpha and TNF cytokines at differen