14-9778-37

antibody from Invitrogen Antibodies

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

Western blot (Recommended by provider)

ELISA (Recommended by provider)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Data presented on provider website)

Flow cytometry (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

14-9778-37

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

2 mg

Concentration

0.5 mg/mL

Storage

4° C

References

Submitted references

BAD-LAMP controls TLR9 trafficking and signalling in human plasmacytoid dendritic cells.

Combes A, Camosseto V, N'Guessan P, Argüello RJ, Mussard J, Caux C, Bendriss-Vermare N, Pierre P, Gatti E
Nature communications 2017 Oct 13;8(1):913

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[Development of EEG changes in subacute leukoencephalitis].

Sofijanov N, Dukovski M, Sadikario A
Godisen zbornik na Medicinskiot fakultet vo Skopje 1977;23:729-38

Other assay

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