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Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Human LAMP1.
(Peptide available as ab25744.)
Our Abpromise guarantee covers the use of ab24170 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IP||Use at an assay dependent concentration. PubMed: 21152024|
|IHC-P||1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ICC||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 1 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 120 kDa).Can be blocked with Human LAMP1 peptide (ab25744).
Abcam recommends using BSA as the blocking agent.
|IHC-Fr||Use at an assay dependent concentration.|
ab24170 staining Lamp-1 in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab24170 at 1μg/ml and ab7291 (staining Tubulin) at 1μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 2 μg/ml (shown in green) and AlexaFluor®594 Goat anti-Mouse (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI, which was added to the secondary antibody mixture.
NOTE: The filamentous pattern observed only occurs when cell are fixed with 100% methanol. We are aware that the staining pattern can change if the cells are fixed with 4%PFA or an alternative fixative. LAMP-1 is a lysosome marker but also plays a role in cellular adhesion. Therefore, lysosomal and filamentous patterns for LAMP-1 can be possible.
ab24170 staining LAMP1 in Human Retinal Microvascular Endothelial Cells (HRMEC) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton in PBS and blocked with 10% serum for 30 minutes at 22°C. Samples were incubated with primary antibody (1/300 in PBS) for 1 hour at 22°C. A diluted (1/500) Alexa Fluor® 488-conjugated Donkey anti-rabbit IgG polyclonal was used as the secondary antibody.
Immunocytochemical immunofluorescence analyis of PFA-fixed rat primary alveolar type II cells, labelling LAMP1 with ab24170 at a dilution of 1/200 incubated for 1/200. Blocking was with 10% BSA incubated for 5 minutes. Secondary was a donkey anti-rabbit polyclonal Alexa Fluor® 647 at 1/400.
This image is courtesy of an anonymous AbreviewBlocking Step: 5% Milk for 1 hour at 20°C
IHC-P image of LAMP1 staining on human Cortex sections using ab24170 (1:400). The sections were deparaffinized and subjected to heat mediated antigen retreival using citric acid. The sections were then permeabilized using 0.05% Tween-20 and blocking was performed using 3% BSA for 1 hour at 21°C. The primary antibody ab24170 was diluted using 3% BSA with 0.05% Tween-20 in PBS and incubated with the sections for 18 hours at 4°C. The secondary antibody used was Goat polyclonal to rabbit IgG conjugated to biotin (1:500)
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