Ubiquilin 2 Antibody (5F5) - Azide and BSA Free

Novus Biologicals | Catalog # H00029978-M03

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, Knockdown Validated

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2a Kappa Clone # 5F5

Format

Azide and BSA Free
View all Ubiquilin 2 Primary Antibodies »

Product Specifications

Immunogen

UBQLN2 (NP_038472, 555 a.a. ~ 624 a.a) partial recombinant protein with GST tag. MW of the GST tag alone is 26 KDa. PNQQFIQQMVQALAGANAPQLPNPEVRFQQQLEQLNAMGFLNREANLQALIATGGDINAAIERLLGSQPS

Specificity

UBQLN2 (5F5)

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2a Kappa

Scientific Data Images for Ubiquilin 2 Antibody (5F5) - Azide and BSA Free

Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

Ubiquilin-2-Antibody-5F5-Western-Blot-H00029978-M03-img0014.jpg
Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

Ubiquilin-2-Antibody-5F5-Western-Blot-H00029978-M03-img0015.jpg
Immunocytochemistry/ Immunofluorescence: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

Immunocytochemistry/ Immunofluorescence: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

Immunocytochemistry/Immunofluorescence: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - Analysis of monoclonal antibody to UBQLN2 on A-431 cell. Antibody concentration 10 ug/ml
Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - Analysis of UBQLN2 expression in PC-12 (Cat # L012V1).
Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - Analysis of UBQLN2 expression in Raw 264.7 (Cat # L024V1).
Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - Analysis of UBQLN2 expression in A-431 (Cat # L015V1).
Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

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.
ELISA: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

ELISA: Ubiquilin 2 Antibody (5F5) [H00029978-M03]

ELISA: Ubiquilin 2 Antibody (5F5) [H00029978-M03] - Detection limit for recombinant GST tagged UBQLN2 is approximately 0.03ng/ml as a capture antibody.
Ubiquilin 2 Antibody (5F5)

Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03] -

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 (https://pubmed.ncbi.nlm.nih.gov/24475300), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Ubiquilin 2 Antibody (5F5)

Western Blot: Ubiquilin 2 Antibody (5F5) [H00029978-M03] -

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 (https://dx.plos.org/10.1371/journal.pone.0136362), licensed under a CC0-1.0 license. Not internally tested by Novus Biologicals.

Applications for Ubiquilin 2 Antibody (5F5) - Azide and BSA Free

Application
Recommended Usage

ELISA

1:100-1:2000

Immunocytochemistry/ Immunofluorescence

1:10-1:500

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Frozen

1:10-1:500

Immunohistochemistry-Paraffin

1:10-1:500
Application Notes
Antibody reactive against cell lysate and recombinant protein for Western Blot. Has also been used for immunofluoresence and ELISA. Immunohistochemistry was reported in scientific literature. Use in Immunohistochemistry-Frozen reported in scientific literature (PMID 24475300)

Reviewed Applications

Read 1 review rated 3 using H00029978-M03 in the following applications:

Formulation, Preparation, and Storage

Purification

IgG purified

Formulation

In 1x PBS, pH 7.4

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

Concentrations vary lot to lot. See vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: Ubiquilin 2

This gene encodes an ubiquitin-like protein (ubiquilin) that shares high degree of similarity with related products in yeast, rat and frog. Ubiquilins contain a N-terminal ubiquitin-like domain and a C-terminal ubiquitin-associated domain. They physically associate with both proteasomes and ubiquitin ligases, and thus thought to functionally link the ubiquitination machinery to the proteasome to affect in vivo protein degradation. This ubiquilin has also been shown to bind the ATPase domain of the Hsp70-like Stch protein.

Alternate Names

Chap1, CHAP1/DSK2, DSK2, DSK2 homolog, 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

Entrez Gene IDs

29978 (Human)

Gene Symbol

UBQLN2

OMIM

300264 (Human)

UniProt

NP_038472 (Human)

Additional Ubiquilin 2 Products

Product Documents for Ubiquilin 2 Antibody (5F5) - Azide and BSA Free

Certificate of Analysis

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Product Specific Notices for Ubiquilin 2 Antibody (5F5) - Azide and BSA Free

This product is produced by and distributed for Abnova, a company based in Taiwan.

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.

Citations for Ubiquilin 2 Antibody (5F5) - Azide and BSA Free

Customer Reviews for Ubiquilin 2 Antibody (5F5) - Azide and BSA Free (1)

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Customer Images

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  • Ubiquilin 2 Antibody (5F5)
    Name: Anonymous
    Application: Western Blot
    Sample Tested: SH-SY5Y cell lysate
    Species: Human
    Verified Customer | Posted 02/24/2017
    Western blot of UBQLN2 shRNA knockdown in SH-SY5Y cells. First lane is the ladder, then treatment with scrambled shRNA and third lane is UBQLN2 knockdown.
    The samples were lysed in Ripa buffer, followed by sonication. 30 ug of the protein sample was loaded on the gel and transferred to the nitrocellulose membrane. Blocked for 45 min at RT in 5 % milk in TBST, overnight incubation in primary antibodies at 4 °C in 5 % milk in TBST, washed with TBST 3x for 10 min, incubated in secondary antibodies (Goat Anti-mouse DyLight 550) at RT for 2 h and then imaged using BioRad ChemiDoc. Images were quantified using ImageLab Software (BioRad). 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, yet if comparing to the data provided in the antibody datasheet, my shRNA wouldn’t really be targeting UBQLN2, but some undefined band. It is possible that the antibody is not able to differentiate between UBQLN2 and the member of the same family, UBQLN1. UBQLN1 is slightly smaller than UBQLN2, but they both share a significant similarity at the C-terminus, from which the immunogen for the UBQLN2 antibody was raised.
    Ubiquilin 2 Antibody (5F5) - Azide and BSA Free H00029978-M03

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Protocols

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FAQs for Ubiquilin 2 Antibody (5F5) - Azide and BSA Free

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  • Q: Do you know if the mAb detects ubiquilin 1?

    A: Based on the sequence homology, it looks like there could be some cross-reactivity with Ubiquilin 1. The immunogen sequence has 92% homology with bovine Ubiquilin 1.

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