Revision 1
#9765
Store at -20C
Vesicle Trafficking Antibody Sampler Kit
1 Kit
(8 x 20 microliters)
877-616-CELL (2355)
877-678-TECH (8324)
3 Trask Lane | Danvers | Massachusetts | 01923 | USA
For Research Use Only. Not for Use in Diagnostic Procedures.
| Product Includes | Product # | Quantity | Mol. Wt | Isotype/Source |
|---|---|---|---|---|
| Phospho-Caveolin-1 (Tyr14) Antibody | 3251 | 20 µl | 23, 25 kDa | Rabbit |
| Caveolin-1 (D46G3) XP® Rabbit mAb | 3267 | 20 µl | 21, 24 kDa | Rabbit IgG |
| Clathrin Heavy Chain (D3C6) XP® Rabbit mAb | 4796 | 20 µl | 190 kDa | Rabbit IgG |
| APPL1 (D83H4) XP® Rabbit mAb | 3858 | 20 µl | 82 kDa | Rabbit IgG |
| EEA1 (C45B10) Rabbit mAb | 3288 | 20 µl | 170 kDa | Rabbit IgG |
| Syntaxin 6 (C34B2) Rabbit mAb | 2869 | 20 µl | 32 kDa | Rabbit IgG |
| Rab5A (E6N8S) Mouse mAb | 46449 | 20 µl | 25 kDa | Mouse IgG1 |
| GOPC (D10A12) Rabbit mAb | 8576 | 20 µl | 59 kDa | Rabbit IgG |
| Anti-rabbit IgG, HRP-linked Antibody | 7074 | 100 µl | Goat | |
| Anti-mouse IgG, HRP-linked Antibody | 7076 | 100 µl | Horse |
Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.
Description
The Vesicle Trafficking Antibody Sampler kit provides an economical means to analyze proteins involved in the intracellular transport of cargo proteins. This kit includes enough primary and secondary antibody to perform two western blot experiments.
Storage
Background
Vesicle trafficking is an integral cellular process and the associated proteins involved also play major roles in other signaling pathways. Caveolins are involved in diverse biological functions including vesicular trafficking, cholesterol homeostasis, cell adhesion, apoptosis, and are also indicated in neurodegenerative disease (1). It is believed that caveolins serve as scaffolding proteins for the integration of signal transduction. Phosphorylation at Tyr14 is essential for caveolin association with SH2 or PTB domain-containing adaptor proteins, such as GRB7 (2-4).
Clathrin-coated vesicles provide for the intracellular transport of proteins following endocytosis and during multiple vesicle trafficking pathways. Vesicles form at specialized areas of the cell membrane where clathrin and associated proteins form clathrin-coated pits. Invagination of these cell membrane-associated pits internalizes proteins and forms an intracellular clathrin-coated vesicle (5,6). Clathrin is the most abundant protein in these vesicles and is present as a basic assembly unit called a triskelion. Each clathrin triskelion is composed of three clathrin heavy chains and three clathrin light chains. Clathrin heavy chain proteins are composed of several functional domains that associate with other vesicle proteins (6).
The APPL1 multidomain adaptor protein is a BAR-domain protein family member that is involved in membrane trafficking within a number of signal transduction pathways (7).
EEA1 is an early endosomal marker and a Rab5 effector protein essential for early endosomal membrane fusion and trafficking (8,9). Syntaxin 6 is a ubiquitously expressed S25C family member of the SNARE proteins (10,11). Syntaxin 6 protein is localized to the trans-Golgi and within endosomes and regulates membrane trafficking by partnering with a variety of other SNARE proteins (12-14). It has two coiled-coil domains (CC1 and CC2) located in the amino-terminal region and a PDZ domain in the carboxy-terminal region (15). The CC2 domain and its adjacent linker region mediate the association of GOPC with the Golgi protein golgin-160 and the Q-SNARE protein syntaxin 6 (15,16). The PDZ domain of GOPC interacts with the carboxy terminus of target proteins to mediate target protein vesicular trafficking and surface expression (17-20).
Rab5 is a member of the Ras superfamily of small Rab GTPases. Rab5 is localized at the plasma membrane and early endosomes and functions as a key regulator of vesicular trafficking during early endocytosis (21).
Background References
- Smart, E.J. et al. (1999) Mol Cell Biol 19, 7289-304.
- Nomura, R. and Fujimoto, T. (1999) Mol Biol Cell 10, 975-86.
- Volonté, D. et al. (2001) J Biol Chem 276, 8094-103.
- Lee, H. et al. (2000) Mol Endocrinol 14, 1750-75.
- Rodriguez-Boulan, E. et al. (2005) Nat Rev Mol Cell Biol 6, 233-47.
- Mousavi, S.A. et al. (2004) Biochem J 377, 1-16.
- Habermann, B. (2004) EMBO Rep 5, 250-5.
- Mu, F.T. et al. (1995) J Biol Chem 270, 13503-11.
- Christoforidis, S. et al. (1999) Nature 397, 621-5.
- Bock, J.B. et al. (2001) Nature 409, 839-41.
- Bock, J.B. et al. (1996) J Biol Chem 271, 17961-5.
- Wendler, F. and Tooze, S. (2001) Traffic 2, 606-11.
- Bock, J.B. et al. (1997) Mol Biol Cell 8, 1261-71.
- Mallard, F. et al. (2002) J Cell Biol 156, 653-64.
- Charest, A. et al. (2001) J Biol Chem 276, 29456-65.
- Hicks, S.W. and Machamer, C.E. (2005) J Biol Chem 280, 28944-51.
- Cheng, J. et al. (2002) J Biol Chem 277, 3520-9.
- He, J. et al. (2004) J Biol Chem 279, 50190-6.
- Wente, W. et al. (2005) J Biol Chem 280, 32419-25.
- Ito, H. et al. (2006) Biochem J 397, 389-98.
- Zerial, M. and McBride, H. (2001) Nat Rev Mol Cell Biol 2, 107-17.
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
Limited Uses
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
Revision 1
Western blot analysis of extracts from various cell lines using Syntaxin 6 (C34B2) Rabbit mAb.
Western blot analysis of extracts from HeLa cells, untreated (-) or H2O2-treated (+), using Phospho-Caveolin-1 (Tyr14) Antibody (upper) and Caveolin-1 (D46G3) XP® Rabbit mAb #3267 (lower).
Western blot analysis of extracts from various cell types using Caveolin-1 (D46G3) XP® Rabbit mAb.
Revision 1
Confocal immunofluorescent analysis of fixed frozen mouse pancreas labeled with Caveolin-1 (D46G3) XP® Rabbit mAb (left, red), and colabeled with DyLight™ 488 Phalloidin #12935 (right, green) and DAPI #4083 (right, blue).
Confocal immunofluorescent analysis of fixed frozen mouse kidney labeled with Caveolin-1 (D46G3) XP® Rabbit mAb (left, red), and colabeled with DyLight™ 488 Phalloidin #12935 (right, green) and DAPI #4083 (right, blue).
Confocal immunofluorescent analysis of fixed frozen mouse skeletal muscle labeled with Caveolin-1 (D46G3) XP® Rabbit mAb (left, red), and colabeled with DyLight™ 488 Phalloidin #12935 (right, green) and DAPI #4083 (right, blue).
Revision 1
Western blot analysis of extracts from various cell lines using EEA1 (C45B10) Rabbit mAb.
Confocal immunofluorescent tile scan of a fixed frozen brain from an amyloid mouse model of Alzheimer's disease using EEA1 (C45B10) Rabbit mAb #3288 (green), GFAP (GA5) Mouse mAb #3670 (red), and DAPI #4083 (blue).
Confocal immunofluorescent analysis of fixed frozen mouse thalamus, labeled with EEA1 (C45B10) Rabbit mAb #3288 (left, green) and co-labeled with GFAP (GA5) Mouse mAb #3670 (right, red) and DAPI #4083 (right, blue).
Revision 1
Confocal immunofluorescent analysis of fixed frozen mouse hippocampus, labeled with EEA1 (C45B10) Rabbit mAb #3288 (left, green) and co-labeled with GFAP (GA5) Mouse mAb #3670 (right, red) and DAPI #4093 (right, blue).
Western blot analysis of extracts from various cell types using APPL1 (D83H4) XP® Rabbit mAb.
Western blot analysis of extracts from various cell lines using Rab5A (E6N8S) Mouse mAb.
Revision 1
Western blot analysis of extracts from various cell lines using Clathrin Heavy Chain (D3C6) XP® Rabbit mAb.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.
Revision 1
Western blot analysis of extracts from various cell lines using GOPC (D10A12) Rabbit mAb.
Confocal immunofluorescent analysis of MCF7 cells using Syntaxin 6 (C34B2) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).
Western blot analysis of extracts from control HeLa cells (lane 1) or caveolin-1 knockout HeLa cells (lane 2) using Caveolin-1 (D46G3) XP® Rabbit mAb #3267 (upper), or β-actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the caveolin-1 knockout HeLa cells confirms the specificity of the antibody to caveolin-1.
Revision 1
Confocal immunofluorescent analysis of HeLa cells using EEA1 (C45B10) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).
Confocal immunofluorescent analysis of HeLa cells using APPL1 (D83H4) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Western blot analysis of HAP1 extracts from WT (left) or RAB5A KO (right) using Rab5A (E6N8S) Mouse mAb. Membranes stained with Ponceau S for total protein normalization (lower). These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies, as a companion to validation data generated by CST scientists.
Revision 1
Confocal immunofluorescent analysis of SH-SY5Y cells using Clathrin Heavy Chain (D3C6) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunoprecipitation of caveolin-1 from HeLa cells using Caveolin-1 (D46G3) XP® Rabbit mAb followed by western blot using the same antibody. Lane 1 is 5% input.
Immunoprecipitation of Rab5A protein from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Mouse (G3A1) mAb IgG1 Isotype Control #5415, lane 3 is Rab5A (E6N8S) Mouse mAb without HeLa cell extracts, and lane 4 is Rab5A (E6N8S) Mouse mAb. Western blot analysis was performed using Rab5A (E6N8S) Mouse mAb.
Revision 1
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Caveolin-1 (D46G3) XP® Rabbit mAb.
Immunoprecipitation of Rab5A protein from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is Rab5A (E6N8S) Mouse mAb. Western blot analysis was performed using Rab5 (C8B1) Rabbit mAb #3547.
Immunohistochemical analysis of paraffin-embedded human lymphoma using Caveolin-1 (D46G3) XP® Rabbit mAb.
Revision 1
Immunohistochemical analysis of paraffin-embedded human ductal carcinoma of the breast using Rab5A (E6N8S) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded mouse lung using Caveolin-1 (D46G3) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using Rab5A (E6N8S) Mouse mAb.
Revision 1
Confocal immunofluorescent analysis of C2C12 cells using Caveolin-1 (D46G3) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using Rab5A (E6N8S) Mouse mAb (left) compared to concentration matched Mouse (G3A1) IgG1 Isotype Control #5415 (right).
Flow cytometric analysis of HeLa cells using Caveolin-1 (D46G3) XP® Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Revision 1
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using Rab5A (E6N8S) Mouse mAb.
Confocal immunofluorescent analysis of HeLa cells using Rab5A (E6N8S) Mouse mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
