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<strong>Figure 1 Western Blot Validation in Human Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: TIP47 3883, (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution
<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Human Cell Lines</strong><br>Loading: 15 µg of lysates per lane.Antibodies: TIP47 3881 (4 μg/mL), TIP47 3883 (1 μg/mL), and beta-actin (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Western Blot Validation in Mouse Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: TIP47 3883 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP
<strong>Figure 4 Western Blot Validation in Mouse EL4 Cells</strong><br>Loading: 15 μg of lysates per lane.Antibodies: TIP47 3883, 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Lane 1: 1 μg/mL Lane 2: 2 μg/mL
<strong>Figure 5 Immunohistochemistry Validation of TIP47 in Rat Liver Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded rat liver tissue using anti-TIP47 antibody (3883) at 2 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
<strong>Figure 6 Induced Expression Validation of TIP47 in Atg7 Liver KO Mice (Singh et al., 2009) </strong><br>TIP47 expression detected by anti-TIP47 antibodies was markedly increased in the liver of Atg7 liver KO mice.
<strong>Figure 7 Induced Expression Validation of TIP47 in Mouse Liver (Singh et al., 2009) </strong><br>TIP47 expression detected by anti-TIP47 antibodies was increased in the autophagic vacuoles (AVs) and lysosomes (Lys) after starvation.
<strong>Figure 8 Induced Expression Validation of TIP47 in Rat Skeletal Muscle (Ramos et al., 2015) </strong><br>Mitochondrial red gastrocnemius muscle PLIN3 (TIP47) content was increased in endurance-trained rats as compared to sedentary rats. PLIN3 protein was detected by anti-TIP47 antibodies.
<strong>Figure 9 Induced Expression Validation of TIP47 in HT29 Cells (Qi et al., 2013) </strong><br>TIP47 expression detected by anti-TIP47 antibodies was increased in the colonic cells after oleic acid treatment.
Figure 1 Western Blot Validation in Human Cell Lines
Loading: 15 μg of lysates per lane.Antibodies: TIP47 3883, (1 µg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution
Cell Treatment
IAV
Most Cited
Trial Size

TIP47 Antibody

CATALOG NUMBER: 3883

Clonality:
Polyclonal
Tested Applications:
ELISA, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Homology:
Predicted species reactivity based on immunogen sequence: Pig: (86%), Rat (79%)
Immunogen:
Anti-TIP47 antibody (3883) was raised against a peptide corresponding to 14 amino acids near the amino terminus of human TIP47.
The immunogen is located within amino acids 30 - 80 of TIP47.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IHC-P, WB
Application Note:
WB: 1-4 μg/mL; IHC-P: 2 μg/mL.
Antibody validated: Western Blot in human and mouse samples; Immunohistochemistry in rat samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1287 - EL4 Cell Lysate
Positive Control 2:
Cat. No. 1464 - Rat Liver Tissue Lysate
Positive Control 3:
Cat. No.1211 - HepG2 Cell Lysate
Positive Control 4:
Cat. No. 1282 – 3T3/NIH Cell Lysate
Predicted Molecular Weight:
Predicted: 47kD and 46kD
Observed: 47kD and 46kD
Validation:

Independent Antibody Validation in Cell lines (Figure 2) shows similar TIP47 expression profile in human cell lines detected by two independent anti-TIP47 antibodies that recognize different epitopes, 3883 against N-terminus domain and 3881 against C-terminus domain.  TIP47 proteins are detected in the most tested cell lines at different expression levels by the two independent antibodies. 

Induced Expression Validation (Figure 6-9): TIP47 expression detected by anit-TIP47 antibodies was increased in Atg7 liver KO mice (figure 6), after starvation (figure 7), after training (figure 8) or after oleic acid treatment (figure 9).

Isoforms:
Human TIP47 has 4 isoforms, including isoform 1 (434aa, 47kD), isoform 2 (251aa, 28kD), isoform 3 (433aa, 47kD), isoform 4 (422aa, 46kD). Mouse TIP47 has one isoform (437aa, 47kD) and rat TIP47 also has one isoform (438aa, 47kD). 3883 can detect human isoforms 1, 3 and 4, and can also detect mouse and rat TIP47.
Purification:
TIP47 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
TIP47 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
TIP47 antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
PLIN3
Additional Names:
TIP47 Antibody: PP17, TIP47, M6PRBP1, Perilipin-3, 47 kDa mannose 6-phosphate receptor-binding protein, 47 kDa MPR-binding protein
Protein Accession Number:
O60664
Protein Gi Number:
68846601
Ncbi Gene Id Number:
10226
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
TIP47 Antibody: Tail-interacting protein (TIP47) is a cytosolic protein essential for the transport of mannose-6-phosphate receptors (MPRs) from endosomes to the trans-Golgi compartments in cells. TIP47 is recruited from the cytoplasm to the late endosomal surface by the Ras-associated protein Rab9 GTPase, enabling it to bind more efficiently to MPR cytoplasmic domains. Recently, it was shown that TIP47 regulates the expression of Rab9, as expression of siRNA to TIP47 in transfected cells dramatically decreased the half-life of the Rab9 protein in addition to stabilizing the subcellular localization of Rab9. At least two isoforms of TIP47 are known to exist.
Background Reference 1:
Barbero et al. J. Cell Biol. 2002; 156:511-8.
Background Reference 2:
Carroll et al. Science 2001; 292:1373-7.
Background Reference 3:
Aivazian et al. J. Cell Biol. 2006; 173:917-26.
Background Reference 4:
Ganley et al. Mol. Biol. Cell 2004; 15:5420-30.
Citation 1:
Singh et al. Autophagy regulates lipid metabolism. Nature. 2009;458(7242):1131-5. PMID: 19339967
Citation 2:
Ramos et al. Changes in mitochondrial perilipin 3 and perilipin 5 protein content in rat skeletal muscle following endurance training and acute stimulated contraction. Exp Physiol. 2015;100(4):450-62. PMID: 25663294
Citation 3:
Qi et al. FOXO3 growth inhibition of colonic cells is dependent on intraepithelial lipid droplet density. J Biol Chem. 2013;288(23):16274-81. PMID: 23603907
Citation 4:
Yamaguchi et al. Characterization of lipid droplets in steroidogenic MLTC-1 Leydig cells: Protein profiles and the morphological change induced by hormone stimulation. Biochim Biophys Acta. 2015;1851(10):1285-95. PMID: 26143378
Citation 5:
Ramos et al. Adipose tissue depot specific differences of PLIN protein content in endurance trained rats. Adipocyte. 2016;5(2):212-23. PMID: 27386161
Citation 6:
Nimura et al. Olanzapine promotes the accumulation of lipid droplets and the expression of multiple perilipins in human adipocytes. Biochem Biophys Res Commun. 2015;467(4):906-12. PMID: 26471304
Citation 7:
Peters et al. Perilipin family (PLIN) proteins in human skeletal muscle: the effect of sex, obesity, and endurance training. Appl Physiol Nutr Metab. 2012;37(4):724-35. PMID: 22667335
Citation 8:
MacPherson et al. A maternal high fat diet has long-lasting effects on skeletal muscle lipid and PLIN protein content in rat offspring at young adulthood. Lipids. 2015;50(2):205-17. PMID: 25552350
Citation 9:
Macpherson et al. Skeletal muscle PLIN3 and PLIN5 are serine phosphorylated at rest and following lipolysis during adrenergic or contractile stimulation. Physiol Rep. 2013;1(4):e00084. PMID: 24303154
Citation 10:
Ramos et al. Higher PLIN5 but not PLIN3 content in isolated skeletal muscle mitochondria following acute in vivo contraction in rat hindlimb. Physiol Rep. 2014;2(10). pii: e12154. PMID: 25318747
Citation 11:
Hunter, BSc. Intra-mitochondrial location of the skeletal muscle perilipin 3 and 5 proteins at rest and following electrically stimulated contraction. Brock University, Canada. 2017.PMID:

FOR RESEARCH USE ONLY

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Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

3883

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

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