The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
Use at an assay dependent concentration.
Use a concentration of 1 µg/ml. Predicted molecular weight: 35 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
LAPTM4B belongs to the LAPTM4/LAPTM5 transporter family. It has an active role in disease progression of malignant cells and is involved in cell proliferation and multidrug resistance. The genetic polymorphism of LAPTM4B is a potential risk factor for the development of colon cancer and gastric cancer. The research results also indicated that LAPTM4B may be a clinically useful prognostic indicator for ovarian carcinoma and may play a role in human hepatocellular carcinoma.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human pineal tissue labelling LAPTM4B with ab82810 at 1/100. A Cy3-conjugated donkey anti-rabbit IgG (1/200) was used as the secondary antibody. Positive staining shown in the membrane and cytoplasm of processes of pinealocytes and intersticial cells. Magnification: 20X. Exposure time: 0.5 - 2.0 seconds. Left - DAPI. Middle - LAPTM4B. Right - Merge.
Western blot - LAPTM4B antibody (ab82810)
Anti-LAPTM4B antibody (ab82810) at 1 µg/ml (in 5% skim milk / PBS buffer) + 293T cell lysate at 10 µg
Secondary HRP conjugated anti-Rabbit IgG at 1/50000 dilution
ICC/IF image of ab82810 stained Hek293 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab82810, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Ndiaye K et al. Differential expression of lysosome-associated protein transmembrane-4 beta (LAPTM4B) in granulosa cells of ovarian follicles and in other bovine tissues. J Ovarian Res8:12 (2015).
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