CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections." alt="ROBO1 was detected in immersion fixed frozen sections of rat embryo (neural tube) using 5 µg/mL Goat Anti-Rat ROBO1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1749) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections." class="big_thumb" />
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | AF1749-SP | |||
HAF007). A specific band was detected for ROBO1 at approximately 220 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and SH-SY5Y human neuroblastoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human ROBO1 Monoclonal Antibody (Catalog # MAB7118) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for ROBO1 at approximately 220 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | MAB7118-SP | |||
HAF016). A specific band was detected for ROBO1 at approximately 200 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of SH-SY5Y human neuroblastoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human ROBO1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7118) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for ROBO1 at approximately 200 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | AF7118-SP | |||
MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B)." alt="HepG2 human hepatocellular carcinoma cell line was stained with Mouse Anti-Human ROBO1 Monoclonal Antibody (Catalog # MAB71181, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B)." class="big_thumb" />
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | MAB71181-SP | |||
| Carrier Free | FAB71181V-100UG | |||
| Carrier Free | FAB71181U-100UG | |||
| Carrier Free | FAB71181T-100UG | |||
| Carrier Free | FAB71181S-100UG | |||
| Carrier Free | FAB71181R-100UG | |||
| Carrier Free | FAB71181N-100UG | |||
| Carrier Free | FAB71181G-100UG | |||
1749-RB) was serially diluted and captured by Mouse Anti-Rat ROBO1 Monoclonal Antibody (Catalog # MAB17491) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Rat ROBO1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1749) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)" alt="Recombinant Rat ROBO1 Fc Chimera (Catalog # 1749-RB) was serially diluted and captured by Mouse Anti-Rat ROBO1 Monoclonal Antibody (Catalog # MAB17491) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Rat ROBO1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1749) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)" class="big_thumb" />
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | MAB17491-SP | |||
| Formulation | Catalog # | Availability | Price | |
|---|---|---|---|---|
| Carrier Free | MAB1749-SP | |||
CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for 
1749-RB) was serially diluted and captured by Mouse Anti-Rat ROBO1 Monoclonal Antibody (Catalog # ![Western Blot: ROBO1 Antibody [NB600-1254] - Western Blot of ROBO1 antibody. Lane M: Super Signal Molecular Weight Marker.Lane 1: HeLa WCL. Load: 30 ug lysate.Primary antibody: ROBO1 antibody at 1:1,000 for overnight at 4C.Secondary antibody: Peroxidase rabbit secondary antibody at 1:40,000 for 30 min at RT.Block: Blocking Buffer for Fluorescent Western Blotting for 30 min at RT.Predicted/Observed size: 181 kDa, 181 kDa for ROBO1. Other band(s): lower bands not identified.](http://images.novusbio.com/fullsize/ROBO1-Antibody-Western-Blot-NB600-1254-img0015.jpg "Western Blot: ROBO1 Antibody [NB600-1254] - Western Blot of ROBO1 antibody. Lane M: Super Signal Molecular Weight Marker.Lane 1: HeLa WCL. Load: 30 ug lysate.Primary antibody: ROBO1 antibody at 1:1,000 for overnight at 4C.Secondary antibody: Peroxidase rabbit secondary antibody at 1:40,000 for 30 min at RT.Block: Blocking Buffer for Fluorescent Western Blotting for 30 min at RT.Predicted/Observed size: 181 kDa, 181 kDa for ROBO1. Other band(s): lower bands not identified.")
![Immunocytochemistry/Immunofluorescence: ROBO1 Antibody [NB600-1254] - Staining of Robo1 in undifferentiated, immortalized human podocytes by Immunocytochemistry/ Immunofluorescence. Cells were fixed with 2% paraformaldehyde and 4% sucrose at room temperature for 10 minutes. The cells were then washed once with PBS, permeabilized with 0.3% Triton X-100 for 10 minutes and incubated with blocking solution (2% FCS, 2% BSA, 0.2% fish gelatin) for 30 minutes, before further incubation with primary Ab for 1 hour. An Alexa Fluor 488 goat anti-rabbit IgG secondary antibody was used at a dilution of 1/200. DAPI was used for nuclear counterstaining.](http://images.novusbio.com/fullsize/ROBO1-Antibody-Immunocytochemistry-Immunofluorescence-NB600-1254-img0005.jpg)
![Immunohistochemistry: ROBO1 Antibody [NB600-1254] - Affinity Purified ROBO1 antibody was used at a concentration of 5 ug/ml to detect ROBO1 in a variety of tissues including multi-human, multi-brain and multi-cancer slides. This image shows staining of human brain tissue. Tissue was formalin-fixed and paraffin embedded.](http://images.novusbio.com/fullsize/ROBO1-Antibody-Immunohistochemistry-NB600-1254-img0016.jpg)
![<b>Biological Strategies Validation. </b>Western Blot: ROBO1 Antibody [NB600-1254] - Biotinylated human mTOR present in a 293T whole cell lysates. Cells were serum-starved for 24 hours prior to harvest. ~20ug of lysate was loaded per lane for SDS-PAGE. Untreated cells are shown in lane 1, whereas cells in lane 2 were treated with IGF-1 (100 ng/ml) for 20 min prior to harvest. Follow reaction of antibody with a 1:2000 dilution of HRP Goat-a-Rabbit IgG for visualization.](http://images.novusbio.com/fullsize/ROBO1-Antibody-Western-Blot-NB600-1254-img0012.jpg)
![Western Blot: ROBO1 Antibody [NB600-1254] - Western blot using Affinity Purified ROBO1 antibody shows detection of a band at ~181 kDa corresponding to ROBO1 present in mouse brain lysate (arrowhead). Approximately 35 ug of lysate was separated by 4-8% SDS-PAGE and transferred onto nitrocellulose. After blocking the membrane was probed with the primary antibody diluted to 1:1,000. Reaction occurred 2h at room temperature followed by washes and reaction with a 1:10,000 dilution of IRDye(TM)800 conjugated Gt-a-Rabbit IgG [H&L] MX for 45 min at room temperature. IRDye(TM)800 fluorescence image was captured using the Odyssey(R) Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.](http://images.novusbio.com/fullsize/ROBO1-Antibody-Western-Blot-NB600-1254-img0013.jpg)
![Western Blot: ROBO1 Antibody [NB600-1254] - Western Blot of ROBO1 antibody. Marker: Opal Pre-stained ladder. Lane 1: HEK293 lysate. Lane 2: HeLa Lysate. Lane 3: MCF-7 Lysate. Lane 4: Jurkat Lysate. Lane 5: A431 Lysate. Lane 6: A549 Lysate. Lane 7: LNCap Lysate. Lane 8: MOLT-4 Lysate. Lane 9: Ramos Lysate. Lane 10: Raji Lysate. Lane 11: A-172 Lysate. Lane 12: NIH/3T3 Lysate. Load: 35 ug per lane. Primary antibody: ROBO1 antibody at 1ug/mL overnight at 4C. Secondary antibody: Peroxidase rabbit secondary antibody at 1:30,000 for 60 min at RT. Blocking Buffer: 1% Casein-TTBS for 30 min at RT.Predicted/Observed size: 181kDa for ROBO1.](http://images.novusbio.com/fullsize/ROBO1-Antibody-Western-Blot-NB600-1254-img0014.jpg)
![Immunohistochemistry: ROBO1 Antibody [NB600-1254] - 1/50 staining mouse lung tissue sections (adult, frozen 100um wholemount sections) by IHC-Fr. The tissue was paraformaldehyde fixed and permeabilized with triton x-100 before incubation with the antibody for 16 hours at 4C.](http://images.novusbio.com/fullsize/ROBO1-Antibody-Immunohistochemistry-NB600-1254-img0011.jpg)
![ELISA: ROBO1 Antibody [NB600-1254] - ELISA results of purified ROBO1 antibody tested against BSA-conjugated peptide of immunizing peptide. Each well was coated in duplicate with 0.1ug of conjugate. The starting dilution of antibody was 5ug/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using 3% fish gel, Goat anti-Rabbit IgG Antibody Peroxidase Conjugated (Min X Bv Ch Gt GP Ham Hs Hu Ms Rt & Sh Serum Proteins) and TMB ELISA Peroxidase Substrate.](http://images.novusbio.com/fullsize/ROBO1-Antibody-ELISA-NB600-1254-img0017.jpg)
NL010) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces and cytoplasm. View our protocol for 
NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces and cytoplasm. View our protocol for ![Western Blot: ROBO1 Antibody (10E2-R) [NBP3-32913] - Western blot analysis of ROBO1 on different lysates with Mouse anti-ROBO1 antibody (NBP3-32913) at 1/1,000 dilution.<br/><br/> Lane 1: MCF-7 cell lysate (10 ug/Lane)<br/> Lane 2: NIH/3T3 cell lysate (10 ug/Lane)<br/> Lane 3: Rat brain tissue lysate (20 ug/Lane)<br/> Lane 4: PC-12 cell lysate (10 ug/Lane)<br/> Lane 5: RAW264.7 cell lysate (10 ug/Lane)<br/><br/> Predicted band size: 181 kDa<br/> Observed band size: 181 kDa<br/><br/> Exposure time: 30 seconds;<br/><br/> 6% SDS-PAGE gel.<br/><br/> Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32913) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1:300,000 dilution was used for 1 hour at room temperature.](http://images.novusbio.com/fullsize/nbp3-32913_mouse-robo1-mab-10e2-r-288202412442967.jpg "Western Blot: ROBO1 Antibody (10E2-R) [NBP3-32913] - Western blot analysis of ROBO1 on different lysates with Mouse anti-ROBO1 antibody (NBP3-32913) at 1/1,000 dilution.<br/><br/> Lane 1: MCF-7 cell lysate (10 ug/Lane)<br/> Lane 2: NIH/3T3 cell lysate (10 ug/Lane)<br/> Lane 3: Rat brain tissue lysate (20 ug/Lane)<br/> Lane 4: PC-12 cell lysate (10 ug/Lane)<br/> Lane 5: RAW264.7 cell lysate (10 ug/Lane)<br/><br/> Predicted band size: 181 kDa<br/> Observed band size: 181 kDa<br/><br/> Exposure time: 30 seconds;<br/><br/> 6% SDS-PAGE gel.<br/><br/> Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32913) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1:300,000 dilution was used for 1 hour at room temperature.")
![Immunohistochemistry: ROBO1 Antibody (10E2-R) [NBP3-32913] - Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-ROBO1 antibody (NBP3-32913) at 1/600 dilution.<br/><br/> The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32913) at 1/600 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.](http://images.novusbio.com/fullsize/nbp3-32913_mouse-robo1-mab-10e2-r-29820241416189.jpg)
![Western blot analysis on different lysates with NBP3-32913 at 1/1,000 dilution. Lane 1: HeLa cell lysate (15 µg/Lane) Lane 2: Rat brain tissue lysate (40 µg/Lane) Predicted band size: 181 kDa Observed band size: 181 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. NBP3-32913 at 1/1,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat anti-Mouse IgG [HRP] at 1/50,000 dilution was used for 1 hour at room temperature.](http://images.novusbio.com/fullsize/nbp3-32913_-western-blot-639204590073436243.jpg)
![Immunocytochemistry/Immunofluorescence: ROBO1 Antibody [NBP3-23373] - Immunocytochemical labeling of ROBO1 in mouse C2C12. The cells were labeled with rabbit polyclonal ROBO1 (C-terminal region) in the absence or presence of blocking peptide. The antibody was then detected using appropriate secondary antibodies conjugated to Cy3.](http://images.novusbio.com/fullsize/nbp3-23373_rabbit-robo1-pab-19920231856944.jpg "Immunocytochemistry/Immunofluorescence: ROBO1 Antibody [NBP3-23373] - Immunocytochemical labeling of ROBO1 in mouse C2C12. The cells were labeled with rabbit polyclonal ROBO1 (C-terminal region) in the absence or presence of blocking peptide. The antibody was then detected using appropriate secondary antibodies conjugated to Cy3.")
![Western Blot: ROBO1 Antibody [NBP3-23373] - Western blot analysis of HepG2 (lanes 1 & 4), C2C12 (lanes 2 & 5), and HUVEC (lanes 3 & 6). The blot was probed with anti-ROBO1 (C-terminal region) in absence (lanes 1-3) or presence of ROBO1 (C-terminal region) blocking peptide (lanes 4-6).](http://images.novusbio.com/fullsize/nbp3-23373_rabbit-robo1-pab-199202317594418.jpg)
![Western Blot: ROBO1 Knockout A549 Cell Lysate [NBP3-18631] - Lane 1: Opal Prestained MW Marker. Lane 2: A549 ROBO KO Clone 5. Lane 3: A549 ROBO KO Clone 15. Lane 4: A549 ROBO KO Clone 17. Lane 5: A549 ROBO KO Clone 18. Lane 6: A549 ROBO KO Clone 21. Lane 7: A549 ROBO1 KO Bulk. Lane 8: A549 WCL Parental. Lane 9: MCF-7 WCL. Load: 35ug lysate/lane. Primary Antibody [Blot A] Anti-ROBO1 ~181kDa; [Blot B] stripped and re-probed with Anti-Tubulin ~50kDa; at 1ug/mL overnight at 2-8C. Secondary Antibody: Goat Anti-Rabbit IgG HRP at 1:70,000 for 30min at RT. Block: TTBS/Casein at RT. No detection of expected band at ~181kDa is observed in ROBO1 knockout A549 when compared with unmodified A549 cell lysates by Western blot.](http://images.novusbio.com/fullsize/ROBO1-Knockout-A549-Cell-Lysate-Western-Blot-NBP3-18631-img0001.jpg "<b>Genetic Strategies Validation. </b>Western Blot: ROBO1 Knockout A549 Cell Lysate [NBP3-18631] - Lane 1: Opal Prestained MW Marker. Lane 2: A549 ROBO KO Clone 5. Lane 3: A549 ROBO KO Clone 15. Lane 4: A549 ROBO KO Clone 17. Lane 5: A549 ROBO KO Clone 18. Lane 6: A549 ROBO KO Clone 21. Lane 7: A549 ROBO1 KO Bulk. Lane 8: A549 WCL Parental. Lane 9: MCF-7 WCL. Load: 35ug lysate/lane. Primary Antibody [Blot A] Anti-ROBO1 ~181kDa; [Blot B] stripped and re-probed with Anti-Tubulin ~50kDa; at 1ug/mL overnight at 2-8C. Secondary Antibody: Goat Anti-Rabbit IgG HRP at 1:70,000 for 30min at RT. Block: TTBS/Casein at RT. No detection of expected band at ~181kDa is observed in ROBO1 knockout A549 when compared with unmodified A549 cell lysates by Western blot.")
