PICALM Signaling Antibody Sampler Kit
Catalog# HAK21026
PICALM Signaling Antibody Sampler Kit
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WB
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IHC-P
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Human
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Mouse
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Rat
General Overview
Kit Components
| Product Includes | Specification | Application | Reactivity | Mw |
|---|---|---|---|---|
| PICALM[HA721706] | 20µl | WB,IHC-P | Human,Mouse,Rat | Predicted band size: 71 kDa |
| Clathrin heavy chain[ET1704-50] | 20µl | WB,IHC-P | Human,Mouse,Rat,Monkey | 180kDa |
| LAMP1[ET1701-94] | 20µl | WB,IHC-P,IF-Tissue | Human | Predicted band size: 45 kDa |
| EEA1[HA722147] | 20µl | WB,IF-Cell,FC | Human,Mouse,Rat,Monkey | Predicted band size: 162 kDa |
| Cathepsin D[ET1608-49] | 20µl | WB,IF-Cell,IHC-P,IP | Human,Mouse,Rat | Predicted band size: 45/28 kDa |
| Goat Anti-Rabbit IgG (H+L)[HA1001] | 100µl | WB,ELISA,IHC-P | Rabbit |
Product Description
Product Features
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Background
The antibodies in this kit serve to characterize phosphatidylinositol-binding clathrin assembly protein (PICALM)-mediated lysosomal maturation, as endo-lysosomal systems are important for normal physiology and prevention of common late-onset neurodegenerative diseases such as Alzheimer's disease (AD). PICALM is a clathrin-binding protein involved in the endo-lysosomal pathway, where it has been genetically associated with AD .</br>PICALM disruption increases the number of early endosomes, which is linked to exacerbated tau aggregation. Early endosome antigen 1 (EEA1) is an early endosomal marker and a Rab5 effector protein essential for early endosomal membrane fusion and trafficking . Lysosome-associated membrane protein 1 (LAMP1) is an abundant lysosomal membrane protein involved in regulating lysosomal motility during lysosome-phagosome fusion . Cathepsin D (CSTD) is a ubiquitously expressed lysosomal aspartyl protease involved in the normal degradation of proteins. Mutations in PICALM were shown to cause lysosomal enzymes and membrane proteins to be mis-trafficked and accumulated.
Data Links
Background References
1. Kaksonen, M. and Roux, A. (2018) Nat Rev Mol Cell Biol 19, 313-326.
2. Hattersley, K.J. et al. (2021) Biochem Biophys Res Commun 570, 103-109.
3. Mu, F.T. et al. (1995) J Biol Chem 270, 13503-11.
4. Christoforidis, S. et al. (1999) Nature 397, 621-5.
5. Eskelinen, E.L. et al. (2003) Trends Cell Biol 13, 137-45.
6. Huynh, K.K. et al. (2007) EMBO J 26, 313-24.
7. Faust, P.L. et al. (1985) Proc Natl Acad Sci USA 82, 4910-4.
Images
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Western blot analysis of PICALM on different lysates with Rabbit anti-PICALM antibody (HA721706) at 1/1,000 dilution.
Lane 1: HepG2 cell lysate
Lane 2: PC-3M cell lysate
Lane 3: A431 cell lysate
Lane 4: K-562 cell lysate
Lane 5: Neuro-2a cell lysate
Lane 6: NIH/3T3 cell lysate
Lane 7: PC-12 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 71 kDa
Observed band size: 65/71 kDa
Exposure time: 3 minutes;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721706) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of Cathepsin D on different lysates with Rabbit anti-Cathepsin D antibody (ET1608-49) at 1/2,000 dilution.
Lane 1: HepG2 cell lysate (15 µg/Lane)
Lane 2: MCF7 cell lysate (15 µg/Lane)
Lane 3: Neuro-2a cell lysate (15 µg/Lane)
Lane 4: Mouse brain tissue lysate (20 µg/Lane)
Predicted band size: 45/28 kDa
Observed band size: 45/28 kDa
Exposure time: 3 minutes;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1608-49) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of Clathrin heavy chain on different lysates with Rabbit anti-Clathrin heavy chain antibody (ET1704-50) at 1/1,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: A431 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: PC-12 cell lysate
Lane 5: HUVEC cell lysate
Lane 6: SH-SY5Y cell lysate
Lane 7: COS-1 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 180kDa
Observed band size: 180 kDa
Exposure time: 10 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1704-50) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of LAMP1 on different lysates with Rabbit anti-LAMP1 antibody (ET1701-94) at 1/1,000 dilution.
Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: Jurkat cell lysate (20 µg/Lane)
Lane 3: HepG2 cell lysate (20 µg/Lane)
Lane 4: HUVEC cell lysate (20 µg/Lane)
Lane 5: Human kidney tissue lysate (40 µg/Lane)
Predicted band size: 45 kDa
Observed band size: 100-120 kDa
Exposure time: 1 minute 2 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-94) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
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