製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to USP3
  • アプリケーション
    適用あり: ICC/IF, WB, ELISAmore details
  • 種交差性
    交差種: Human
    交差が予測される動物種: Mouse, Rat, Rabbit, Horse, Chicken, Guinea pig, Cow, Cat, Dog, Pig, Zebrafish
  • 免疫原

    Synthetic peptide corresponding to a region within the internal sequence amino acids 180 - 229 (IEQFCCYFKE LPAVELRNGK TAGRRTYHTR SQGDNNVSLV EEFRKTLCAL) of Human USP3 (NP_006528)

  • ポジティブ・コントロール
    • Jurkat cell lysate. This antibody gave a positive result when used in the following methanol fixed cell lines: HeLa.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab82935 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 59 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
ELISA Use at an assay dependent concentration.

Titre using peptide based assay 1/12500.

ターゲット情報

  • 機能
    Hydrolase that deubiquitinates monoubiquitinated target proteins such as histone H2A and H2B. Required for proper progression through S phase and subsequent mitotic entry. May regulate the DNA damage response (DDR) checkpoint through deubiquitination of H2A at DNA damage sites. Associates with the chromatin.
  • 組織特異性
    Expressed in all tissues examined, with strongest expression in pancreas.
  • 配列類似性
    Belongs to the peptidase C19 family. USP3 subfamily.
    Contains 1 UBP-type zinc finger.
  • ドメイン
    Both protease activity and an intact zinc finger are required for H2A monodeubiquitination.
  • 細胞内局在
    Nucleus. Localizes preferentially with monoubiquitinated H2A to chromatin.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Deubiquitinating enzyme 3 antibody
    • EC 3.1.2.15 antibody
    • SIH003 antibody
    • Ubiquitin carboxyl-terminal hydrolase 3 antibody
    • Ubiquitin specific peptidase 3 antibody
    • Ubiquitin specific processing protease 3 antibody
    • Ubiquitin Specific Protease 3 antibody
    • Ubiquitin thioesterase 3 antibody
    • Ubiquitin thiolesterase 3 antibody
    • Ubiquitin-specific-processing protease 3 antibody
    • UBP antibody
    • UBP3_HUMAN antibody
    • Usp3 antibody
    see all

画像

  • ICC/IF image of ab82935 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab82935 at 5µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • Anti-USP3 antibody (ab82935) at 1 µg/ml + Jurkat cell lysate at 10 µg

    Secondary
    HRP conjugated anti-Rabbit IgG at 1/50000 dilution

    Predicted band size : 59 kDa
    Observed band size : 65 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 38 kDa. We are unsure as to the identity of these extra bands.Gel concentration 12%

参考文献

ab82935 has not yet been referenced specifically in any publications.

レビューと Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% TritonX100 in PBS
Specification
HeLa
Fixative
Paraformaldehyde
Username

Dr. Kirk Mcmanus

Verified customer

投稿 Aug 12 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (U2OS)
Loading amount
20 µg
Specification
U2OS
Treatment
0.1 uM Doxorubicin for 3 hours
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Dr. Andrew Cobb

Verified customer

投稿 Nov 07 2012

Thank you for your reply.

Either of these antibodies should work well in your assay. Ab101473 has already been tested with HeLa lysates and works well, however I know that the exposure time was an important variable in your decision. Since th...

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Thank you again for your patience.

I have heard back from the lab that this antibody was also tested with 721_B lymphoblasts, which have been shown to express USP3.

I hope this information will be useful, but please let me know if y...

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Thank you for your call last week and for your patience whileI have been in touch with the lab.

The exposure time used to create this image was 1 minute. We are lookingthrough our files to see if the antibody has been tested with other cell l...

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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