![Western Blot: RGL2 Antibody (4D10) [H00005863-M02] - Impact of RGL2 depletion on RalB activation at autophagosomes upon starvation. Validation of RGL2 depletions. Representative western blots for RGL2 and GAPDH from cell lysates of HEK-HT cells prepared 72 hrs after transfection with the indicated siRNAs. Quantifications of RGL2 protein depletion, normalized for siControl condition (=100), are shown below the WB. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-019-45443-1), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/RGL2-Antibody-4D10-Western-Blot-H00005863-M02-img0009.jpg "Western Blot: RGL2 Antibody (4D10) [H00005863-M02] - Impact of RGL2 depletion on RalB activation at autophagosomes upon starvation. Validation of RGL2 depletions. Representative western blots for RGL2 and GAPDH from cell lysates of HEK-HT cells prepared 72 hrs after transfection with the indicated siRNAs. Quantifications of RGL2 protein depletion, normalized for siControl condition (=100), are shown below the WB. Image collected and cropped by CiteAb from the following publication (//www.nature.com/articles/s41598-019-45443-1), licensed under a CC-BY license.")
| Reactivity | HuSpecies Glossary |
| Applications | WB, ELISA, ICC/IF |
| Clone | 4D10 |
| Clonality | Monoclonal |
| Host | Mouse |
| Conjugate | Unconjugated |
| Description | Quality control test: Antibody Reactive Against Recombinant Protein. |
| Immunogen | RGL2 (AAH32681, 644 a.a. ~ 743 a.a) partial recombinant protein with GST tag. MW of the GST tag alone is 26 KDa. PGASDCRIIRVQMELGEDGSVYKSILVTSQDKAPSVISRVLKKNNRDSAVASEYELVQLLPGERELTIPASANVFYAMDGASHDFLLRQRRRSSTATPGV |
| Specificity | RGL2 - ral guanine nucleotide dissociation stimulator-like 2 |
| Isotype | IgG1 |
| Clonality | Monoclonal |
| Host | Mouse |
| Gene | RGL2 |
| Purity | IgG purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
| Dilutions |
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| Application Notes | Antibody reactivity against cell lysate and recombinant protein for WB. It has also been used for IF and ELISA. |
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| Publications |
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| Storage | Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles. |
| Buffer | In 1x PBS, pH 7.4 |
| Preservative | No Preservative |
| Purity | IgG purified |
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Secondary Antibodies |
Isotype Controls |
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![Immunocytochemistry/Immunofluorescence: RGL2 Antibody (4D10) [H00005863-M02] - Analysis of monoclonal antibody to RGL2 on HeLa cell. Antibody concentration 10 ug/ml.](http://images.novusbio.com/fullsize/RGL2-Antibody-4D10-Immunocytochemistry-Immunofluorescence-H00005863-M02-img0005.jpg "Immunocytochemistry/Immunofluorescence: RGL2 Antibody (4D10) [H00005863-M02] - Analysis of monoclonal antibody to RGL2 on HeLa cell. Antibody concentration 10 ug/ml.")
![Western Blot: RGL2 Antibody (4D10) [H00005863-M02] - RGL2 monoclonal antibody (M02), clone 4D10 Analysis of RGL2 expression in HeLa.](http://images.novusbio.com/fullsize/RGL2-Antibody-4D10-Western-Blot-H00005863-M02-img0007.jpg "Western Blot: RGL2 Antibody (4D10) [H00005863-M02] - RGL2 monoclonal antibody (M02), clone 4D10 Analysis of RGL2 expression in HeLa.")
![ELISA: RGL2 Antibody (4D10) [H00005863-M02] - Detection limit for recombinant GST tagged RGL2 is approximately 1ng/ml as a capture antibody.](http://images.novusbio.com/fullsize/RGL2-Antibody-4D10-ELISA-H00005863-M02-img0008.jpg "ELISA: RGL2 Antibody (4D10) [H00005863-M02] - Detection limit for recombinant GST tagged RGL2 is approximately 1ng/ml as a capture antibody.")
![Immunocytochemistry/ Immunofluorescence: RGL2 Antibody (4D10) [H00005863-M02] - Localization of RGL2 at different endomembrane compartments. HEK-HT (Normal cells) & HEK-HT-H-RasV12 (Transformed cells) cells were fixed & imaged for endogenous RGL2 (IF anti-RGL2, represented in red), together with EEA1 (early endosome marker, IF anti-EEA1, represented in green), GFP-Rab11 (recycling endosome marker, represented in green), GFP-Rab6 (Trans-Golgi marker, represented in green), or iRFP-LC3 (autophagosome marker, represented in green). Representative confocal cross sections of normal cells are shown (left). Quantifications are reported for both normal & transformed cells (right). Localization was calculated as percentage (%) of the indicated endomembrane compartments positive for RGL2, as compared to the control pseudo-compartments. Each dot corresponds to one cell. Graph represents mean ± SEM of 21 to 40 cells from 3–4 independent experiments. For statistics Mann Whitney test was used. **p value < 0.01, ***p value < 0.001. Scale bars are 10 µm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31222145), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/h00005863-m02_mouse-monoclonal-rgl2-antibody-4d10-310202415482283.jpg "Immunocytochemistry/ Immunofluorescence: RGL2 Antibody (4D10) [H00005863-M02] - Localization of RGL2 at different endomembrane compartments. HEK-HT (Normal cells) & HEK-HT-H-RasV12 (Transformed cells) cells were fixed & imaged for endogenous RGL2 (IF anti-RGL2, represented in red), together with EEA1 (early endosome marker, IF anti-EEA1, represented in green), GFP-Rab11 (recycling endosome marker, represented in green), GFP-Rab6 (Trans-Golgi marker, represented in green), or iRFP-LC3 (autophagosome marker, represented in green). Representative confocal cross sections of normal cells are shown (left). Quantifications are reported for both normal & transformed cells (right). Localization was calculated as percentage (%) of the indicated endomembrane compartments positive for RGL2, as compared to the control pseudo-compartments. Each dot corresponds to one cell. Graph represents mean ± SEM of 21 to 40 cells from 3–4 independent experiments. For statistics Mann Whitney test was used. **p value < 0.01, ***p value < 0.001. Scale bars are 10 µm. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31222145), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
![Western Blot: RGL2 Antibody (4D10) [H00005863-M02] - Impact of RGL2 depletion on RalB activation at autophagosomes upon starvation. (A) Modulation by autophagy of RalB activity at autophagosomes upon RGL2 depletion. Normal HEK-HT cells were transfected with indicated siRNAs (day 0), then transfected with FRET RalB biosensor & Cherry-LC3 plasmids (day 2), & incubated in either basal (DMEM) or starvation (EBSS) medium (4 hours) before imaging (day 3). Graph represents mean ± SEM of n = 17 to 57 cells per condition, from 3 independent experiments. To compare DMEM versus EBSS conditions Mann Whitney test was used, **p < 0.01, ***, & p < 0.001. (B) Validation of RGL2 depletions. Representative western blots for RGL2 & GAPDH from cell lysates of HEK-HT cells prepared 72 hrs after transfection with the indicated siRNAs. Quantifications of RGL2 protein depletion, normalized for siControl condition (=100), are shown below the WB. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31222145), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/h00005863-m02_mouse-monoclonal-rgl2-antibody-4d10-310202415485145.jpg "Western Blot: RGL2 Antibody (4D10) [H00005863-M02] - Impact of RGL2 depletion on RalB activation at autophagosomes upon starvation. (A) Modulation by autophagy of RalB activity at autophagosomes upon RGL2 depletion. Normal HEK-HT cells were transfected with indicated siRNAs (day 0), then transfected with FRET RalB biosensor & Cherry-LC3 plasmids (day 2), & incubated in either basal (DMEM) or starvation (EBSS) medium (4 hours) before imaging (day 3). Graph represents mean ± SEM of n = 17 to 57 cells per condition, from 3 independent experiments. To compare DMEM versus EBSS conditions Mann Whitney test was used, **p < 0.01, ***, & p < 0.001. (B) Validation of RGL2 depletions. Representative western blots for RGL2 & GAPDH from cell lysates of HEK-HT cells prepared 72 hrs after transfection with the indicated siRNAs. Quantifications of RGL2 protein depletion, normalized for siControl condition (=100), are shown below the WB. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/31222145), licensed under a CC-BY license. Not internally tested by Novus Biologicals.")
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![SDS-Page: Mouse IgG1 Isotype Control (MG1) [NBP1-97005] - Lane 1: , Non-reduced. M: Opal Pre-stained Ladder. Lane 2: , Reduced. Load: 1.0 ug per lane. Predicted/Observed: 120 kDa Non-reduced, 55 and 25 Reduced.](https://images.novusbio.com/images/Mouse-IgG1-Isotype-Control-MG1-SDS-Page-NBP1-97005-img0002.jpg "SDS-Page: Mouse IgG1 Isotype Control (MG1) [NBP1-97005] - Lane 1: , Non-reduced. M: Opal Pre-stained Ladder. Lane 2: , Reduced. Load: 1.0 ug per lane. Predicted/Observed: 120 kDa Non-reduced, 55 and 25 Reduced.")