Ubiquilin 2 Antibody (6H9) - BSA Free

Immunocytochemistry/ Immunofluorescence: 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.

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.

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.

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.

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.

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.

Western Blot: 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).

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).

Immunocytochemistry/ Immunofluorescence: 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).

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, ICC/IF, IHC, IP, KO, Mycoplasma
• Clone: 6H9
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

Ubiquilin 2 Antibody (6H9) - BSA Free Summary

• Description: Novus Biologicals Knockout (KO) Validated Mouse Ubiquilin 2 Antibody (6H9) - BSA Free (NBP2-25164) is a monoclonal antibody validated for use in IHC, WB, ICC/IF, Simple Western and IP. Anti-Ubiquilin 2 Antibody: Cited in 17 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: Human Ubiquilin 2 expressed in and purified from E. coli [UniProt# Q9UHD9]
• Localization: Cytoplasm. Nucleus.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: UBQLN2
• Purity: Protein G purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:1000
  • Immunohistochemistry 1:1000
  • Immunohistochemistry-Frozen
  • Immunoprecipitation
  • Knockdown Validated
  • Knockout Validated
  • Simple Western 1:100
  • Western Blot 1:1000-1:2000
• Application Notes: This Ubiquilin 2 (6H9). antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry, and Western Blot, where a band can be seen at approx. 66-68 kDa.
  
  In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
  See Simple Western Antibody Database for Simple Western validation: Tested in HeLa lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:100, apparent MW was 74 kDa. Use in Immunohistochemistry-Frozen reported in scientific literature (PMID:33028421).
• Theoretical MW: 66-68 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: 50% PBS, 50% glycerol
• Preservative: 5mM Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein G purified

Alternate Names for Ubiquilin 2 Antibody (6H9) - BSA Free

• Chap1
• CHAP1/DSK2
• DSK2 homolog
• DSK2
• FLJ10167
• FLJ56541
• hPLIC-2
• N4BP4LIC-2
• Nedd4 binding protein 4
• PLIC-2
• PLIC2Dsk2
• Protein linking IAP with cytoskeleton 2
• RIHFB2157
• ubiquilin 2
• ubiquilin-2
• Ubiquitin-like product Chap1/Dsk2

Background

Ubiquilin 2, also known as PLIC2 and Chap1, is a member of the ubiquilin protein family, which regulate the degradation of cellular proteins through proteasome or autophage-like pathways. Humans have four ubiquilin genes, each encoding a separate protein referred to as Ubiquilin 1, 2, 3 and 4. All ubiquilins contain an N-terminal ubiquitin-like (UBL) domain and a C-terminal ubiquitin-associated (UBA) domain, while the central part of the molecules are highly variable. The UBL domains bind subunits of the proteasome, and the UBA domains binds to polyubiquitin chains that are typically conjugated onto proteins marked for proteosomal degradation. Ubiquilin 2 has a unique region close to the C terminus containing 12 PXX tandem collagen like repeats, where P is proline and X is most cases valine, glycine, isoleucine or threonine. Teepu Siddique and his collaborators have identified mutations in the ubiquilin 2 gene leading to protein point mutations which were important contributors to several forms of amyotrophic lateral sclerosis (ALS) and Frontotemporal lobar degeneration (FTLD). Interestingly, these mutations involved alterations in proline residues in the PXX repeat region (P497H, P497S, P506T, P509S and P525S). Recently, the Lee and Trojanowski group investigated C9orf72 hexanucleotide expansion and ubiquilin 2 pathology in patients with ALS and FTLD by genetic analysis and immunohistochemistry and found distinct Ubiquilin 2 pathology in ALS and FTLD-TDP with C9orf72 expansion. C9orf72 hexonucleotide expansion is the most common cause to date of familial ALS and FTLD. Ubiquilin 2 protein is of different molecular size in mouse and human, 638 and 624 amino acids respectively. As a result the mouse protein, endogenously expressed in rodent 3T3 cells, runs on SDS-PAGE and western blots slightly slower than the human protein.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Ubiquilin 2 Antibody (NBP2-25164)(21)

Publications using NBP2-25164

• McDowell, I;Ayoubi, R;Fotouhi, M;Southern, K;McPherson, PS;Laflamme, C;NeuroSGC/YCharOS/EDDU collaborative group, ;ABIF Consortium, ; The identification of high-preforming antibodies for Ubiquilin-2 for use in Western Blot, immunoprecipitation, and immunofluorescence F1000Research 2023-03-30 [PMID: 37359784] (Immunocytochemistry/ Immunofluorescence, Mouse)
• Nementzik L, Thumbadoo K, Murray H et al. Distribution of ubiquilin 2 and TDP-43 aggregates throughout the CNS inUBQLN2p.T487I-linked amyotrophic lateral sclerosis and frontotemporal dementia bioRxiv 2023-02-10 (IHC, Human)
• Takei T, Sasazawa Y, Noshiro D et al. Ubiquilin-2 liquid droplets catalyze alpha -synuclein fibril formation. The EMBO Journal 2025-10-14 [PMID: 41087580]
• Almaguer-Mederos L, Kandi A, Sen N et al. Spinal cord phosphoproteome of SCA2 mouse model reveals alteration of ATXN2-N-term PRM-SH3-actin interactome and of autophagy. Molecular & Cellular Proteomics : MCP 2025-09-22 [PMID: 40992657]
• Mohan, HM;Fernandez, MG;Huang, C;Lin, R;Ryou, JH;Seyfried, D;Grotewold, N;Barget, AJ;Whiteley, AM;Basrur, V;Mosalaganti, S;Paulson, HL;Sharkey, LM; Endogenous retrovirus-like proteins recruit UBQLN2 to stress granules and shape their functional biology Science advances 2025-07-18 [PMID: 40680123]
• Laura R. Nementzik, Kyrah M. Thumbadoo, Helen C. Murray, David Gordon, Shu Yang, Ian P. Blair, Clinton Turner, Richard L. M. Faull, Maurice A. Curtis, Catriona McLean, Garth A. Nicholson, Molly E. V. Swanson, Emma L. Scotter Distribution of ubiquilin 2 and TDP ‐43 aggregates throughout the CNS in UBQLN2 p. T487I ‐linked amyotrophic lateral sclerosis and frontotemporal dementia Brain Pathology 2023-12-19 [PMID: 38115557]
• Dao TP, Rajendran A, Galagedera SKK et al. Short disordered N-termini & proline-rich domain are major regulators of UBQLN1/2/4 phase separation Biophysical journal 2023-11-30 [PMID: 38041404] (WB)
• Dao TP, Rajendran A, Galagedera SKK et al. Short N-terminal disordered regions and the proline-rich domain are major regulators of phase transitions for full-length UBQLN1, UBQLN2 and UBQLN4 bioRxiv : the preprint server for biology 2023-09-29 [PMID: 37808720] (WB)
• Buel GR, Chen X, Myint W et al. E6AP AZUL interaction with UBQLN1/2 in cells, condensates, and an AlphaFold-NMR integrated structure Structure (London, England : 1993) 2023-02-14 [PMID: 36827983]
• Phung TH, Tatman M, Monteiro MJ UBQLN2 undergoes a reversible temperature-induced conformational switch that regulates binding with HSPA1B: ALS/FTD mutations cripple the switch but do not destroy HSPA1B binding Biochimica et biophysica acta. General subjects 2022-11-21 [PMID: 36423739] (WB, Human)
• Mohan HM, Trzeciakiewicz H, Pithadia A et al. RTL8 promotes nuclear localization of UBQLN2 to subnuclear compartments associated with protein quality control Cellular and molecular life sciences : CMLS 2022-03-05 [PMID: 35247097] (ICC/IF, WB, Human)
• Lin BC, Phung TH, Higgins NR et al. ALS/FTD mutations in UBQLN2 are linked to mitochondrial dysfunction through loss-of-function in mitochondrial protein import Human molecular genetics 2021-04-22 [PMID: 33891006]
• Higgins NR, Greenslade JE, Wu JJ et al. Serpin neuropathology in the P497S UBQLN2 mouse model of ALS/FTD Brain pathology (Zurich, Switzerland) 2021-03-29 [PMID: 33780087] (WB, IF/IHC, Mouse)
• Gerson JE, Linton H, Xing J, et al. Shared and divergent phase separation and aggregation properties of brain-expressed ubiquilins Scientific reports 2021-01-11 [PMID: 33431932] (WB, Human)
• Whiteley AM, Prado MA, de Poot SAH et Al. Global proteomics of Ubqln2-based murine models of ALS J Biol Chem 2020-12-10 [PMID: 33277362] (Western Blot)
• Wang S, Tatman M, Monteiro MJ Overexpression of UBQLN1 reduces neuropathology in the P497S UBQLN2 mouse model of ALS/FTD Acta Neuropathol Commun 2020-10-07 [PMID: 33028421] (IHC-Fr, Mouse)
• Alexander EJ, Ghanbari Niaki A, Zhang T et al. Ubiquilin 2 modulates ALS/FTD-linked FUS-RNA complex dynamics and stress granule formation. Proc. Natl. Acad. Sci. U.S.A. 2018-11-15 [PMID: 30442662] (WB, Human)
• Huang N, Erie C, Lu ML, Wei J. Aberrant subcellular localization of SQSTM1/p62 contributes to increased vulnerability to proteotoxic stress recovery in Huntington's disease. Mol. Cell. Neurosci. 2017-12-12 [PMID: 29246658] (Mouse)
• Teyssou E, Chartier L, Amador MD et al. Novel UBQLN2 mutations linked to amyotrophic lateral sclerosis and atypical hereditary spastic paraplegia phenotype through defective HSP70-mediated proteolysis Neurobiology of Aging 2017-10-01 [PMID: 28716533] (Immunocytochemistry/ Immunofluorescence, Mouse)
• Le NT, Chang L, Kovlyagina I et al. Motor neuron disease, TDP-43 pathology, and memory deficits in mice expressing ALS-FTD-linked UBQLN2 mutations. Proc. Natl. Acad. Sci. U.S.A. 2016-11-22 [PMID: 27834214]
• Hjerpe R, Bett JS, Keuss MJ et al. UBQLN2 Mediates Autophagy-Independent Protein Aggregate Clearance by the Proteasome. Cell. 2016-08-11 [PMID: 27477512] (ICC/IF, Mouse)

Reviews for Ubiquilin 2 Antibody (NBP2-25164) (2)

Average Rating: 5

(Based on 2 reviews)

We have 2 reviews tested in 1 species: Human.

reviewed by: Verified Customer IP Human 08/19/2022

Summary

• Application: Immunoprecipitation
• Sample Tested: 293 whole cell lysate
• Species: Human
• Lot: 082019

Comments

• Comments: Antibody was used at 1:30 dilution and was crosslinked to Protein A/G beads. HEK293 cells were lysed in 1x Tris buffered saline containing 1% NP40 and 5% glycerol. 600 micrograms of whole cell lysate was used for IP. IP was done for 2 hours.

reviewed by: Verified Customer ICC Human 02/09/2022

Summary

• Application: Immunocytochemistry
• Sample Tested: U2OS cells
• Species: Human
• Lot: 6H9

Comments

• Comments: Dilution: 1:1000

Bioinformatics

• Gene Symbol: UBQLN2
• Entrez:
  • 29978(Human)
• OMIM:
  • 300264(Human)
• Uniprot:
  • Q9UHD9(Human)