TIP47 Antibody

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ProSci
Product Code: PSI-3883
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-3883-0.02mg0.02mg£150.00
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PSI-3883-0.1mg0.1mg£449.00
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Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunohistochemistry (IHC)
  • Western Blot (WB)

Images

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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 ug/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution
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<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Human Cell Lines</strong><br> Loading: 15 ug 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.
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<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
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<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
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<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.
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<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.
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<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.
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<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.
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<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.
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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 ug/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 ug 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.

Documents

Further Information

Additional Names:
TIP47 Antibody: PP17, TIP47, M6PRBP1, Perilipin-3, 47 kDa mannose 6-phosphate receptor-binding protein, 47 kDa MPR-binding protein
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.
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 References:
  • Barbero et al. J. Cell Biol. 2002; 156:511-8.
  • Carroll et al. Science 2001; 292:1373-7.
  • Aivazian et al. J. Cell Biol. 2006; 173:917-26.
  • Ganley et al. Mol. Biol. Cell 2004; 15:5420-30.
Buffer:
TIP47 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Conjugate:
Unconjugated
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.
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.
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.
NCBI Gene ID #:
10226
NCBI Official Name:
perilipin 3
NCBI Official Symbol:
PLIN3
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 47kD and 46kD

Observed: 47kD and 46kD
Protein Accession #:
O60664
Protein GI Number:
68846601
Purification:
TIP47 Antibody is affinity chromatography purified via peptide column.
Research Area:
Signal Transduction
Swissprot #:
O60664
User NOte:
Optimal dilutions for each application to be determined by the researcher.
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).

References

  1. Singh et al. Autophagy regulates lipid metabolism. Nature. 2009;458(7242):1131-5. PMID: 19339967
  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
  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
  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
  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
  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
  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
  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
  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
  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
  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:

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