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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, M | WB, ICC | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MABC83 |
| Description | Anti-Vps4/SKD1 Antibody, clone 5E6.1 |
| Alternate Names |
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| Background Information | Vacuolar protein sorting-associated protein 4B (Vps4), or SKD1, is a ubiquitous AAA ATPase involved in endosomal vesicle trafficking. Trafficking of vesicles containing proteins such as receptors, from the endosome to the cytoplasm, is an important regulatory mechanism for cell-surface receptors and the signaling pathways that involve those receptors. Such endosomal vesicles are shuttled from the endosome to the cytoplasm by multivesicular bodies (MVB). MVBs are large multi-protein complexes that assemble and disassemble during this process. Previous studies have indicated that the yeast homologue of Vps4, Vps4p (60 % sequence identity to Vps4), interacts with endosomes via its N-terminal microtubule interacting and trafficking (MIT) domain, and also associates with a number of MVB proteins including Vps2p, Bro1p, Vps20p, Vta1p, and Did2p/Chm1p. Although the precise role of Vps4 is still unclear, previous studies have suggested that it interacts with the ESCRT-III complex involved in the sorting of MVBs and has been associated with virus-loaded vesicles such those in HIV-infected cells. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | Purified mouse monoclonal IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mm NaCl with 0.05% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-Vps4/SKD1 Antibody, clone 5E6.1 is an antibody against Vsp4/SKD1 for use in Western Blotting, ICC. |
| Key Applications |
|
| Application Notes | Immunocytochemistry Analysis: A 1:500 dilution from a representative lot detected Vps4/SKD1 in NIH/3T3, A431, and HeLa cells. |
| Biological Information | |
|---|---|
| Immunogen | Linear peptide corresponding to human Vps4/SKD1. |
| Clone | 5E6.1 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Isotype | IgG1κ |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | The protein encoded by this gene is a member of the AAA protein family (ATPases associated with diverse cellular activities), and is the homolog of the yeast Vps4 protein. In humans, two paralogs of the yeast protein have been identified. The former share a high degree of aa sequence similarity with each other, and also with yeast Vps4 and mouse Skd1 proteins. Mouse Skd1 (suppressor of K+ transport defect 1) has been shown to be a yeast Vps4 ortholog. Functional studies indicate that both human paralogs associate with the endosomal compartments, and are involved in intracellular protein trafficking, similar to Vps4 protein in yeast. The gene encoding this paralog has been mapped to chromosome 18; the gene for the other resides on chromosome 16. |
| Gene Symbol |
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| Purification Method | Protein g Purified |
| UniProt Number |
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| UniProt Summary | FUNCTION: Involved in late steps of the endosomal multivesicular bodies (MVB) pathway. Recognizes membrane-associated ESCRT-III assemblies and catalyzes their disassembly, possibly in combination with membrane fission. Redistributes the ESCRT-III components to the cytoplasm for further rounds of MVB sorting. MVBs contain intraluminal vesicles (ILVs) that are generated by invagination and scission from the limiting membrane of the endosome and mostly are delivered to lysosomes enabling degradation of membrane proteins, such as stimulated growth factor receptors, lysosomal enzymes and lipids. In conjunction with the ESCRT machinery also appears to function in topologically equivalent membrane fission events, such as the terminal stages of cytokinesis and enveloped virus budding (HIV-1 and other lentiviruses). SUBUNIT STRUCTURE: Proposed to be monomeric or homodimeric in nucleotide-free form and to oligomerize upon binding to ATP to form two stacked hexameric or heptameric rings with a central pore through which ESCRT-III substrates are translocated in an ATP-dependent manner. In vitro, associates on the inside of a helical tubular structure formed by a CHMP2A-CHMP3 polymer. Interacts with CHMP1A, CHMP1B, CHMP2A, CHMP4B and CHMP6. Interacts with VPS4A; the interaction suggests a heteromeric assembly with VPS4A. Interacts with VTA1. SUBCELLULAR LOCATION: Prevacuolar compartment membrane; Peripheral membrane protein. Late endosome membrane; Peripheral membrane protein probable. Note: Membrane-associated in the prevacuolar endosomal compartment. Localized in HIV-1 particles purified from acutely infected cells. TISSUE SPECIFICITY: Ubiquitously expressed. DOMAIN: The MIT domain serves as an adapter for ESCRT-III proteins. It forms an asymmetric three-helix bundle that binds amphipathic MIM (MIT interacting motif) helices along the groove between MIT helices 2 and 3 present in a subset of ESCRT-III proteins thus establishing the canonical MIM-MIT interaction. In an extended conformation along the groove between helices 1 and 3, also binds to a type-2 MIT interacting motif (MIM2). POST-TRANSLATIONAL MODIFICATION: Phosphorylated upon DNA damage, probably by ATM or ATR. SEQUENCE SIMILARITIES: Belongs to the AAA ATPase family. Contains 1 MIT domain. |
| Molecular Weight | ~52 kDa observed. The calculated molecular weight of this protein is ~50 kDa. Uniprot describes that this protein is autophosphoralated upon DNA damage. |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Evaluated by Western Blot in human skeletal muscle tissue lysate. Western Blot Analysis: 1 µg/mL of this antibody detected Vps4/SKD1 in 10 µg of human skeletal muscle tissue lysate. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at 2-8°C from date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |