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![Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - Generation of transgenic mice that overexpress human ubiquilin-1. Immunoblots of equal amounts of total brain lysates from 12 month-old WT mouse, 12 month-old Ubqln-1 48 transgenic mouse, 12 month-old Ubqln-1 62 transgenic mouse and end stage 15 week-old R6/2 transgenic mouse. The top panel was probed with a monoclonal anti-ubiquilin antibody (Invitrogen antibody clone 3D5E2) and the lower panel with a different monoclonal anti-Ubiquilin 2 antibody (Novus antibody clone 5F5). Note two immunoreactive ubiquilin bands are seen at 70 kDa and at 90 kDa, which we presume is a modified form of ubiquilin. Both blots were also probed for actin to ensure equal loading. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0087513) licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-5F5-Western-Blot-H00029978-M03-img0014.jpg "Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - Generation of transgenic mice that overexpress human ubiquilin-1. Immunoblots of equal amounts of total brain lysates from 12 month-old WT mouse, 12 month-old Ubqln-1 48 transgenic mouse, 12 month-old Ubqln-1 62 transgenic mouse and end stage 15 week-old R6/2 transgenic mouse. The top panel was probed with a monoclonal anti-ubiquilin antibody (Invitrogen antibody clone 3D5E2) and the lower panel with a different monoclonal anti-Ubiquilin 2 antibody (Novus antibody clone 5F5). Note two immunoreactive ubiquilin bands are seen at 70 kDa and at 90 kDa, which we presume is a modified form of ubiquilin. Both blots were also probed for actin to ensure equal loading. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0087513) licensed under a CC-BY license.")
![Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - Confirmation of polyQ-associated proteins from PC-12 cells identified by TAPI. Western blot analysis of TAPI-purified polyQ aggregates from PC-12 cells confirms the presence of several disease-associated proteins only in the Htt-Q74 samples. All proteins migrated near their predicted molecular weights. For control, the TAPI procedure was conducted in parallel on the induced Htt-Q23 cell line (FUS, TDP-43, UBQLN2, HNRNPA1) or the un-induced Htt-Q74 cell line (CLINT1, HSPA8, RAD23B, SGTA). Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0136362), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-5F5-Western-Blot-H00029978-M03-img0015.jpg)
![Immunocytochemistry/Immunofluorescence: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - Analysis of monoclonal antibody to UBQLN2 on A-431 cell. Antibody concentration 10 ug/ml](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-5F5-Immunocytochemistry-Immunofluorescence-H00029978-M03-img0007.jpg)
![Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - Analysis of UBQLN2 expression in PC-12 (Cat # L012V1).](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-5F5-Western-Blot-H00029978-M03-img0008.jpg)
![Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - Analysis of UBQLN2 expression in Raw 264.7 (Cat # L024V1).](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-5F5-Western-Blot-H00029978-M03-img0009.jpg)
![Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - Analysis of UBQLN2 expression in A-431 (Cat # L015V1).](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-5F5-Western-Blot-H00029978-M03-img0010.jpg)
![<b>Genetic Strategies Validation. </b>Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - UBQLN2 shRNA knockdown in SH-SY5Y cells. First lane is the ladder, then treatment with scrambled shRNA and third lane is UBQLN2 knockdown. 30 ug of the protein sample was loaded on the gel and transferred to the nitrocellulose membrane. Overnight incubation in primary antibodies at 4 degrees C in 5 % milk in TBST. Then incubated in secondary antibodies (Goat Anti-mouse DyLight 550) at RT for 2 h, and then imaged. Knockdown of the upper band with shRNA for UBQLN2 was about 82 %, and of the lower band only about 8 %, suggesting a high specificity of the used shRNA. This image was submitted via customer Review.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-5F5-Western-Blot-H00029978-M03-img0012.jpg)
![ELISA: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - Detection limit for recombinant GST tagged UBQLN2 is approximately 0.03ng/ml as a capture antibody.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-5F5-ELISA-H00029978-M03-img0006.jpg)
![Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - Generation of transgenic mice that overexpress human ubiquilin-1.(A) Schematic of the Thy1.2 expression construct used to generate ubiquilin-1 transgenic mice. Human ubiquilin-1 with an N-terminal FLAG tag was cloned in the appropriate orientation between the XhoI site of the Thy1.2 expression cassette. (B) Southern Blot of the first generation offspring of two founder mice (48 & 62). (C) Validation of a PCR genotyping protocol. Amplification of the transgene was only observed in mice that Southern blotting revealed to be positive. (D) Immunoblots of brain cortical lysates with an anti-FLAG antibody & for tubulin indicated that line 62 offspring express higher levels of FLAG-ubiquilin-1 than line 48. (E) Immunoblots of equal amounts of total brain lysates from 12 month-old WT mouse, 12 month-old Ubqln-1 48 transgenic mouse, 12 month-old Ubqln-1 62 transgenic mouse & end stage 15 week-old R6/2 transgenic mouse. The top panel was probed with a monoclonal anti-ubiquilin antibody (Invitrogen antibody clone 3D5E2) & the lower panel with a different monoclonal anti-ubiquilin antibody (Novus antibody clone 5F5). Note two immunoreactive ubiquilin bands are seen at ∼70 kDa & at ∼90 kDa, which we presume is a modified form of ubiquilin. Both blots were also probed for actin to ensure equal loading. (F) Cryostat sections of a Ubqln-1 62 transgenic mouse brain (a–f) & WT mouse brain (g-i) showing anti-FLAG antibody staining (Alexa 594, left panels) & corresponding DAPI staining (center panels) & the result of merging the fluorescent & DAPI signals (right hand panels). The brain sections shown are of the hippocampus (a–c & g–i) & cerebellum (d–f). Identical exposure settings were used for the left hand panels. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24475300), licensed under a CC-BY license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/h00029978-m03_mouse-monoclonal-ubiquilin-2-antibody-5f5-310202415482227.jpg)
![Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - Confirmation of polyQ-associated proteins from PC-12 cells identified by TAPI.(A) Western blotting shows that the addition of doxycycline to the PC-12 cell model induces the expression of HttQ74-GFP, resulting in aggregates that can be purified by TAPI. The kinase ERK is probed as a negative control; ERK was never identified by mass spectrometry, so is not expected to co-fractionate with polyQ aggregates. (B) Western blot analysis of TAPI-purified polyQ aggregates from PC-12 cells confirms the presence of several disease-associated proteins only in the Htt-Q74 samples. All proteins migrated near their predicted molecular weights. For control, the TAPI procedure was conducted in parallel on the induced Htt-Q23 cell line (FUS, TDP-43, UBQLN2, HNRNPA1) or the un-induced Htt-Q74 cell line (CLINT1, HSPA8, RAD23B, SGTA). (C) Confocal microscopy shows localization of identified proteins to Htt-Q74 aggregates in PC12 cells. (left) RAD23B, nominally a DNA repair protein, localizes to nuclear Htt-Q74 inclusions but not cytoplasmic inclusions. (middle) FUS, an RNA-binding protein localizes to nuclear & cytoplasmic Htt-Q74 inclusions. (right) CLINT1, a clatherin-interacting protein, is observed in cytoplasmic Htt-Q74 aggregates. Arrows indicate foci with co-localized proteins. Green = GFP; Magenta = CLINT1, FUS or RAD23B in merge. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0136362), licensed under a CC0-1.0 license. Not internally tested by Novus Biologicals.](http://images.novusbio.com/fullsize/h00029978-m03_mouse-monoclonal-ubiquilin-2-antibody-5f5-310202415483625.jpg)
![Knockdown Validated: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - Validation of UBQLN2 antibody for staining of endogenous UBQLN2 in U2OS cells. U2OS cells were transfected with control siRNA or siRNA targeting UBQLN2. Cells were either seeded as separate groups (i.e., control and UBQLN2 siRNA) or mixed 1:1 and seeded together. Cells were stained using the mouse monoclonal anti-UBQLN2 6H9 (Novus NBP2-25164), at 1:250 in 2% BSA PBS for 1h. Secondary antibody was goat Anti-Mouse DyLight 488. Knockdown of UBQLN2 can be clearly seen to decrease the signal, indicating that the antibody is specific to UBQLN2. (G) displays zoom of the indicated areas in. For the mixed cells a white arrowhead indicates a cell transfected with control siRNA and a black arrowhead a cell transfected with UBQLN2 siRNA. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1016/j.cell.2016.07.001), licensed under a CC-BY license.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Knockdown-Validated-NBP2-25164-img0010.jpg "<b>Genetic Strategies Validation. </b>Knockdown Validated: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - Validation of UBQLN2 antibody for staining of endogenous UBQLN2 in U2OS cells. U2OS cells were transfected with control siRNA or siRNA targeting UBQLN2. Cells were either seeded as separate groups (i.e., control and UBQLN2 siRNA) or mixed 1:1 and seeded together. Cells were stained using the mouse monoclonal anti-UBQLN2 6H9 (Novus NBP2-25164), at 1:250 in 2% BSA PBS for 1h. Secondary antibody was goat Anti-Mouse DyLight 488. Knockdown of UBQLN2 can be clearly seen to decrease the signal, indicating that the antibody is specific to UBQLN2. (G) displays zoom of the indicated areas in. For the mixed cells a white arrowhead indicates a cell transfected with control siRNA and a black arrowhead a cell transfected with UBQLN2 siRNA. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1016/j.cell.2016.07.001), licensed under a CC-BY license.")
![<b>Genetic Strategies Validation. </b>Simple Western: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - Simple Western lane view shows a specific band for Ubiquilin 2 in 0.5 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Simple-Western-NBP2-25164-img0007.jpg)
![Western Blot: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - Analysis of different tissue and cell lysates using mouse mAb to ubiquilin 2, NBP2-25164, dilution 1:1,000 in green: [1] protein standard (red), [2] NIH-3T3, [3] C6, [4] HEK293, [5] HeLa, [6] SH-SY5Y, [7] rat whole brain, and [8] mouse whole brain. The band at 65-70kDa corresponds to ubiquilin 2 protein, which is known to differ between the human and rodent proteins.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Western-Blot-NBP2-25164-img0009.jpg)
![Immunocytochemistry/Immunofluorescence: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - Analysis of an NIH-3T3 cell culture stained with mouse mAb to ubiquilin 2, NBP2-25164, dilution 1:1,000 in green, and costained with chicken pAb to lamin A/C, dilution 1:5,000 in red. The blue is DAPI staining of nuclear DNA. The cells were treated with 50uM of chloroquine, an inhibitor of autophagy, for 16 hours prior to staining. NBP2-25164 antibody reveals vesicular staining of ubiquilin 2 protein accumulated in lysosomes in the cytoplasm, while the lamin A/C antibody stains the nuclear lamina.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Immunocytochemistry-Immunofluorescence-NBP2-25164-img0008.jpg)
![Immunocytochemistry/Immunofluorescence: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - Analysis of Human U2OS cells stained with Ubiquilin 2 antibody. Primary antibody dilution: 1:1000. Image from verified customer review.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Immunocytochemistry-Immunofluorescence-NBP2-25164-img0011.jpg)
![Immunocytochemistry/Immunofluorescence: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - HeLa cell cultures stained with NBP2-25164 (green) and chicken polyclonal antibody to Vimentin: NB300-223 (red). In most individual cells Ubiquilin 2 is present diffusely in the cytoplasm of cells, though some cells show enrichment of the protein in spherical autophagosome-like structure.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Immunocytochemistry-Immunofluorescence-NBP2-25164-img0003.jpg)
![<b>Genetic Strategies Validation. </b>Knockout Validated: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - Lysates of HAP1 WT and Ubiquilin 2 KO were prepared, and 30 ug of protein were processed for immunoblot with NBP2-25164. The Ponceau stained transfer of the blot is shown. Antibody dilution used: 1/2000. Predicted band size: 65 kDa. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Knockout-Validated-NBP2-25164-img0012.jpg)
![Immunoprecipitation: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - HAP1 lysates were prepared, and immunoprecipitation was performed using 2.0 ug of Ubiquilin 2 antibody (NBP2-25164) pre-coupled to Dynabeads protein G. Samples were washed and processed for immunoblot with NBP2-25164. For immunoblot, Recombinant Ubiquilin 2 (Rb x Ubiquilin 2) was used at 1/1000. The Ponceau stained transfer of the blot is shown. SM=4% starting material; UB=4% unbound fraction. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Immunoprecipitation-NBP2-25164-img0013.jpg)
![<b>Genetic Strategies Validation. </b>Knockout Validated: Ubiquilin 2 Antibody (6H9) [NBP2-25164] - HAP1 WT and Ubiquilin 2 KO cells were labeled with a green or a far-red fluorescent dye, respectively. WT and KO cells were mixed and plated to a 1:1 ratio in a 96-well plate with glass bottom. Cells were stained with Ubiquilin-2 antibody (NBP2-25164) and with the corresponding Alexa-fluor 555 coupled secondary antibody including DAPI. Acquisition of the blue (nucleus-DAPI), green (identification of WT cells), red (antibody staining) and far-red (identification of KO cells) channels was performed. Representative image of the blue and red (grayscale) channels is shown. WT and KO cells are outlined with green and magenta dashed line, respectively. Antibody dilution used: 1/1000. Bars = 10 um Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-6H9-Knockout-Validated-NBP2-25164-img0014.jpg)
![Western Blot: Ubiquilin 2 Antibody [NBP1-85639] - Analysis in A-431 cells transfected with control siRNA, target specific siRNA probe #1, using Anti-UBQLN2 antibody. Remaining relative intensity is presented. Loading control: Anti-GAPDH.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-Western-Blot-NBP1-85639-img0012.jpg "<b>Genetic Strategies Validation. </b>Western Blot: Ubiquilin 2 Antibody [NBP1-85639] - Analysis in A-431 cells transfected with control siRNA, target specific siRNA probe #1, using Anti-UBQLN2 antibody. Remaining relative intensity is presented. Loading control: Anti-GAPDH.")
![<b>Genetic Strategies Validation. </b>Immunocytochemistry/Immunofluorescence: Ubiquilin 2 Antibody [NBP1-85639] - Staining of human cell line U-2 OS shows localization to plasma membrane & cytosol. Antibody staining is shown in green.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-Immunocytochemistry-Immunofluorescence-NBP1-85639-img0010.jpg)
![Immunohistochemistry-Paraffin: Ubiquilin 2 Antibody [NBP1-85639] - Staining of human cerebellum shows moderate to strong cytoplasmic and nuclear positivity in Purkinje cells.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-Immunohistochemistry-Paraffin-NBP1-85639-img0021.jpg)
![Immunohistochemistry-Paraffin: Ubiquilin 2 Antibody [NBP1-85639] - Staining of human fallopian tube shows weak to moderate cytoplasmic positivity in glandular cells.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-Immunohistochemistry-Paraffin-NBP1-85639-img0019.jpg)
![Immunohistochemistry-Paraffin: Ubiquilin 2 Antibody [NBP1-85639] - Staining of human cerebral cortex shows moderate to strong cytoplasmic positivity in neurons and glial cells.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-Immunohistochemistry-Paraffin-NBP1-85639-img0020.jpg)
![Immunohistochemistry-Paraffin: Ubiquilin 2 Antibody [NBP1-85639] - Staining of human placenta shows moderate to strong cytoplasmic positivity in cytotrophoblast.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-Immunohistochemistry-Paraffin-NBP1-85639-img0022.jpg)
![Simple Western: Ubiquilin 2 Antibody [NBP1-85639] - Simple Western lane view shows a specific band for Ubiquilin 2 in 0.2 mg/ml of RT-4 (Left) and U-251MG (Right) lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-Simple-Western-NBP1-85639-img0006.jpg)
![Simple Western: Ubiquilin 2 Antibody [NBP1-85639] - Electropherogram image(s) of corresponding Simple Western lane view. Ubiquilin 2 antibody was used at 1:20 dilution on RT-4 and U-251MG lysate(s).](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-Simple-Western-NBP1-85639-img0007.jpg)
![Western Blot: Ubiquilin 2 Antibody [NBP2-93303] - Analysis of extracts of various cell lines, using Ubiquilin 2 at 1:1000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit . Exposure time: 90s.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-Western-Blot-NBP2-93303-img0003.jpg "Western Blot: Ubiquilin 2 Antibody [NBP2-93303] - Analysis of extracts of various cell lines, using Ubiquilin 2 at 1:1000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit . Exposure time: 90s.")
![Immunocytochemistry/Immunofluorescence: Ubiquilin 2 Antibody [NBP2-93303] - Analysis of NIH-3T3 cells using Ubiquilin 2 . Blue: DAPI for nuclear staining.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-93303-img0002.jpg)
![Immunohistochemistry-Paraffin: Ubiquilin 2 Antibody [NBP2-93303] - Mouse kidney using UBQLN2 Rabbit pAb at dilution of 1:250 (40x lens).](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-Immunohistochemistry-Paraffin-NBP2-93303-img0005.jpg)
![Immunocytochemistry/Immunofluorescence: Ubiquilin 2 Antibody [NBP2-93303] - Analysis of HeLa cells using Ubiquilin 2 . Blue: DAPI for nuclear staining.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-Immunocytochemistry-Immunofluorescence-NBP2-93303-img0001.jpg)
![Immunohistochemistry-Paraffin: Ubiquilin 2 Antibody [NBP2-93303] - Human esophageal cancer using UBQLN2 Rabbit pAb at dilution of 1:250 (40x lens).](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-Immunohistochemistry-Paraffin-NBP2-93303-img0004.jpg)
![Western Blot: Ubiquilin 2 Antibody [NBP1-79753] - Hela, Antibody Dilution: 1.0 ug/ml UBQLN2 is supported by BioGPS gene expression data to be expressed in HeLa.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-Western-Blot-NBP1-79753-img0004.jpg "Western Blot: Ubiquilin 2 Antibody [NBP1-79753] - Hela, Antibody Dilution: 1.0 ug/ml UBQLN2 is supported by BioGPS gene expression data to be expressed in HeLa.")
![Western Blot: Ubiquilin 2 Antibody [NBP1-79753] - HepG2 cell lysate, concentration 0.2-1 ug/ml.](http://images.novusbio.com/fullsize/Ubiquilin-2-Antibody-Western-Blot-NBP1-79753-img0003.jpg)
![ELISA: Human Ubiquilin 2 - Ready-To-Use ELISA Kit (Colorimetric) [NBP3-38773] - Standard Curve Reference](http://images.novusbio.com/fullsize/nbp3-38773_human-ubiquilin-2-ready-to-use-elisa-kit-colorimetric-2110202415295682.png "ELISA: Human Ubiquilin 2 - Ready-To-Use ELISA Kit (Colorimetric) [NBP3-38773] - Standard Curve Reference")
![ELISA: Human Ubiquilin 2 ELISA Kit (Colorimetric) [NBP3-38774] - Standard Curve Reference](http://images.novusbio.com/fullsize/nbp3-38774_human-ubiquilin-2-elisa-kit-colorimetric-2110202415403188.png "ELISA: Human Ubiquilin 2 ELISA Kit (Colorimetric) [NBP3-38774] - Standard Curve Reference")