Robo1 Antibody [A23H22] | Primary Antibodies

Application: Reactivity: Mouse, Rat, Human

Usage Information

Dilution
WB1:1000 IP1:30 FCM1:500

Application
WB, IP, FCM

Reactivity
Mouse, Rat, Human

Source
Rabbit 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
181 kDa 38 kDa, 120-130 kDa, 215,250 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	Mouse E14.5 brain tissue; C57 E18 brain tissue; Rat E14 brain tissue; U-87 MG cells; HepG2 cells; HeLa cells; HAP1 cells
Negative Control	LoVo cells; MCF7 cells

Datasheet & SDS

Biological Description

Specificity
Robo1 Antibody [A23H22] detects endogenous levels of total Robo1 protein.

Clone
A23H22

Synonym(s)
DUTT1; ROBO1; Roundabout homolog 1; Deleted in U twenty twenty; H-Robo-1

Background
Robo1, or Roundabout guidance receptor 1, also known as DUTT1, is the founding member of the Robo family of single-pass transmembrane glycoproteins, mediating Slit-dependent repulsion that is critical for midline axon guidance, neuronal migration, and tissue morphogenesis. Its extracellular domain comprises five immunoglobulin-like domains Ig1 to Ig5, with Ig1 and Ig2 forming the primary Slit2-D2 binding site and assembling into compact dimers or tetramers via Ig4 contacts, further rigidified by heparan sulfate binding on Ig1 and Ig2. This is followed by three cysteine-rich fibronectin type III domains, Fn1 to Fn3, with Fn2 and Fn3 near the membrane for cell surface positioning, a transmembrane domain, and an intracellular domain of about 150 amino acids containing four conserved CC0 to CC3 motifs. The CC2 and CC3 motifs recruit Abelson kinase and Dock for cytoskeletal repulsion, while CC1 mediates LogR and DCC antagonism. Robo1's core function centers on Slit2-induced signaling, where the ternary Slit2N-Ig1-heparan sulfate complex triggers Robo1 oligomerization without major conformational shift, and the cytoplasmic CC motifs recruit Dock and ELMO, leading to Rac1 and CDC42 activation, Arp2/3 and WAVE complex engagement, and branched F-actin-mediated repulsion at growth cones and leading edges, thereby inhibiting Netrin and DCC attraction and establishing anterior-posterior boundaries. Robo1 patterns commissural axons by facilitating the anterior turn after midline crossing, restricts glial cells to the midline, governs vascular branching in the lung and kidney, and regulates tumor cell migration, with redundancy among Robo1, Robo2, Robo3, and Robo4 ensuring developmental robustness. Epigenetic silencing through hypermethylation or mutations in Robo1 promotes metastasis in breast and lung cancers by increasing invasion, advances glioma progression, and contributes to autism through neural circuit defects, while reactivation of the Slit2-Robo1 pathway restores tumor suppression.

Background

Robo1, or Roundabout guidance receptor 1, also known as DUTT1, is the founding member of the Robo family of single-pass transmembrane glycoproteins, mediating Slit-dependent repulsion that is critical for midline axon guidance, neuronal migration, and tissue morphogenesis. Its extracellular domain comprises five immunoglobulin-like domains Ig1 to Ig5, with Ig1 and Ig2 forming the primary Slit2-D2 binding site and assembling into compact dimers or tetramers via Ig4 contacts, further rigidified by heparan sulfate binding on Ig1 and Ig2. This is followed by three cysteine-rich fibronectin type III domains, Fn1 to Fn3, with Fn2 and Fn3 near the membrane for cell surface positioning, a transmembrane domain, and an intracellular domain of about 150 amino acids containing four conserved CC0 to CC3 motifs. The CC2 and CC3 motifs recruit Abelson kinase and Dock for cytoskeletal repulsion, while CC1 mediates LogR and DCC antagonism. Robo1's core function centers on Slit2-induced signaling, where the ternary Slit2N-Ig1-heparan sulfate complex triggers Robo1 oligomerization without major conformational shift, and the cytoplasmic CC motifs recruit Dock and ELMO, leading to Rac1 and CDC42 activation, Arp2/3 and WAVE complex engagement, and branched F-actin-mediated repulsion at growth cones and leading edges, thereby inhibiting Netrin and DCC attraction and establishing anterior-posterior boundaries. Robo1 patterns commissural axons by facilitating the anterior turn after midline crossing, restricts glial cells to the midline, governs vascular branching in the lung and kidney, and regulates tumor cell migration, with redundancy among Robo1, Robo2, Robo3, and Robo4 ensuring developmental robustness. Epigenetic silencing through hypermethylation or mutations in Robo1 promotes metastasis in breast and lung cancers by increasing invasion, advances glioma progression, and contributes to autism through neural circuit defects, while reactivation of the Slit2-Robo1 pathway restores tumor suppression.

References
https://pubmed.ncbi.nlm.nih.gov/29307485/ https://pubmed.ncbi.nlm.nih.gov/36042257/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

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

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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%