REEP5 Antibody (Mouse mAb) [E6M2]

Application: Reactivity: Human, Pig

Usage Information

Dilution
WB1:1000-1:5000 IHC1:5000-1:20000 IF1:400-1:1600

Application
WB, IHC, IF, ELISA

Reactivity
Human, Pig

Source
Mouse Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW Observed MW
21 kDa 18 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight. Post-translational modifications(e.g., phosphorylation, glycosylation); Splice variants and isoforms; Relative charge; Multimerization.

Positive Control	Pig heart tissue; Human colon cancer tissue; A549 cells; HEK-293 cells; UNCaP cells; HeLa cells; Jurkat cells; MCLT-4 cells; K-562 cells
Negative Control

Datasheet & SDS

Biological Description

Specificity
REEP5 Antibody (Mouse mAb) [E6M2] detects endogenous levels of total REEP5 protein.

Clone
E6M2

Synonym(s)
Receptor expression-enhancing protein 5, Polyposis locus protein 1, Protein TB2, REEP5, C5orf18, DP1, TB2

Background
REEP5, also known as DP1, belongs to the reticulon and DP1/REEP family of membrane-curvature-generating proteins that shape the tubular network of the endoplasmic reticulum, and it shares its highest sequence and functional conservation with the yeast protein Yop1p, which has served as the primary structural model for the family. REEP5 carries a reticulon homology domain built from hydrophobic transmembrane segments long enough to fully traverse the ER membrane, arranged in an unusual hairpin topology in which both the N- and C-termini face the same side of the membrane rather than crossing it in a conventional multi-pass configuration, and this hairpin architecture wedges into the outer leaflet of the bilayer to impose local membrane curvature. Structural characterization of Yop1p identifies a previously uncharacterized amphipathic helix positioned C-terminal to the transmembrane hairpin, and this amphipathic helix is highly conserved in its physicochemical properties and position across both the DP1/REEP and reticulon families; deleting or mutating this helix abolishes the protein's ability to generate membrane tubules, directly establishing the amphipathic helix as functionally required for curvature generation rather than a passive structural extension. REEP5 purifies as a stable homodimer, and biochemical reconstitution shows that REEP proteins use both their transmembrane segments and their amphipathic helix together to introduce high membrane curvature into lipid bilayers, with sufficiently strong curvature-generating activity capable of driving formation of lipoprotein-like particles from reconstituted membranes; synthetic peptides corresponding to the wild-type amphipathic helix sequence disrupt pre-formed ER tubular networks generated in cell-free extract, while peptides carrying point mutations or D-amino acid substitutions in this helix fail to produce the same disruptive effect, confirming that the native, correctly folded amphipathic helix sequence is specifically responsible for its membrane-remodeling activity. Site-directed cross-linking analysis of the transmembrane segments shows that individual transmembrane helices mediate homotypic dimerization independently, allowing the protein to assemble into a curved oligomeric scaffold, and this oligomeric self-association, rather than the intrinsic wedge shape of a single monomer, is the dominant mechanism by which REEP-family proteins stabilize the high curvature characteristic of ER tubules. REEP5 interacts with the reticulon proteins, which are similarly restricted to tubular ER domains and excluded from ER sheets and the nuclear envelope, and together this reticulon-REEP interaction stabilizes tubule morphology through combined wedging and oligomerization. Mutations affecting the equivalent oligomerization interfaces and amphipathic helix in the related paralog REEP1 disrupt ER membrane integrity and are linked to hereditary spastic paraplegia and distal hereditary motor neuropathy.

Background

REEP5, also known as DP1, belongs to the reticulon and DP1/REEP family of membrane-curvature-generating proteins that shape the tubular network of the endoplasmic reticulum, and it shares its highest sequence and functional conservation with the yeast protein Yop1p, which has served as the primary structural model for the family. REEP5 carries a reticulon homology domain built from hydrophobic transmembrane segments long enough to fully traverse the ER membrane, arranged in an unusual hairpin topology in which both the N- and C-termini face the same side of the membrane rather than crossing it in a conventional multi-pass configuration, and this hairpin architecture wedges into the outer leaflet of the bilayer to impose local membrane curvature. Structural characterization of Yop1p identifies a previously uncharacterized amphipathic helix positioned C-terminal to the transmembrane hairpin, and this amphipathic helix is highly conserved in its physicochemical properties and position across both the DP1/REEP and reticulon families; deleting or mutating this helix abolishes the protein's ability to generate membrane tubules, directly establishing the amphipathic helix as functionally required for curvature generation rather than a passive structural extension. REEP5 purifies as a stable homodimer, and biochemical reconstitution shows that REEP proteins use both their transmembrane segments and their amphipathic helix together to introduce high membrane curvature into lipid bilayers, with sufficiently strong curvature-generating activity capable of driving formation of lipoprotein-like particles from reconstituted membranes; synthetic peptides corresponding to the wild-type amphipathic helix sequence disrupt pre-formed ER tubular networks generated in cell-free extract, while peptides carrying point mutations or D-amino acid substitutions in this helix fail to produce the same disruptive effect, confirming that the native, correctly folded amphipathic helix sequence is specifically responsible for its membrane-remodeling activity. Site-directed cross-linking analysis of the transmembrane segments shows that individual transmembrane helices mediate homotypic dimerization independently, allowing the protein to assemble into a curved oligomeric scaffold, and this oligomeric self-association, rather than the intrinsic wedge shape of a single monomer, is the dominant mechanism by which REEP-family proteins stabilize the high curvature characteristic of ER tubules. REEP5 interacts with the reticulon proteins, which are similarly restricted to tubular ER domains and excluded from ER sheets and the nuclear envelope, and together this reticulon-REEP interaction stabilizes tubule morphology through combined wedging and oligomerization. Mutations affecting the equivalent oligomerization interfaces and amphipathic helix in the related paralog REEP1 disrupt ER membrane integrity and are linked to hereditary spastic paraplegia and distal hereditary motor neuropathy.

References
https://pubmed.ncbi.nlm.nih.gov/25646439/ https://pubmed.ncbi.nlm.nih.gov/33495454/

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

Tech Support

Handling Instructions

Tel: +1-832-582-8158 Ext:3

If you have any other enquiries, please leave a message.

Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%