Anti-Ubiquitin抗体[Ubi-1]
参阅全部 Ubiquitin 一抗
小鼠单克隆抗体[Ubi-1] to Ubiquitin
Mouse
适用于: IHC-P, WBmore details
不适用于: IP
与反应: Mouse, Human
Full length protein corresponding to Cow Ubiquitin conjugated to keyhole limpet haemocyanin (Glutaraldehyde).
H9C2 cells IHC-P:FFPE mouse brain alzheimer
This product was changed from ascites to tissue culture supernatant on 17th May 2016. The following lots are from ascites and are still in stock as of 17th May 2016: GR159581-2, GR159581-12, GR159581-13. Lot numbers higher than GR159581-13 will be from tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly.
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Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Preservative: 0.1% Sodium azide
浓度
50 µl 浓度为 1 mg/ml
Affinity purified
单克隆
Ubi-1
IgG1
kappa
Abpromise™承诺保证使用ab7254于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
---|---|---|
IHC-P | Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. | |
WB | (4) | 1/100 - 1/5000. |
应用说明
Is unsuitable for IP.
Entrez Gene: 7314 Human
Entrez Gene: 22187 Mouse
SwissProt: P0CG47 Human
SwissProt: P0CG48 Human
SwissProt: P62979 Human
SwissProt: P62987 Human
SwissProt: P62988 Human
SwissProt: P0CG49 Mouse
SwissProt: P62991 Mouse
Unigene: 356190 Human
Unigene: 520348 Human
Unigene: 282093 Mouse
Unigene: 371592 Mouse
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [Ubi-1] (ab7254)Image from Ohmi K et al., Plos One. 2011;6(11):e27461. Fig 7.; doi: 10.1371/journal.pone.0027461 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Immunohistochemical staining of ubiquitin in a 7 months-old MPS IIIA mouse brain (PFA fixed) using ab7254.
Sections, 40 µm thick, were cut sagittally on a vibratome. Slices were permeabilized in 1% triton X-100/PBS for 1 hour at room temperature. The samples were placed on vectabond-coated glass slides and dried overnight. They were then rehydrated in PBS and washed in ice-cold methanol for 10 min.
The slices were washed in 0.3% H2O2 in methanol at room temperature and incubated sequentially with 0.5% normal donkey serum, ab7254 and biotin-labeled F(ab')2 donkey secondary antibody in Tris-buffered saline (pH 7.5) containing 2% BSA and 0.02% Tween 20. The signal was detected with an ABC kit, visualized with diaminobenzidine and counterstained with hematoxylin.
Western blot - Anti-Ubiquitin antibody [Ubi-1] (ab7254)
All lanes : Anti-Ubiquitin antibody [Ubi-1] (ab7254) at 1/100 dilution
Lane 1 : HeLa cell lysate
Lane 2 : HeLa cell lysate with MG132
Lane 3 : Mouse brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Anti mouse IgG at 1/10000 dilution
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [Ubi-1] (ab7254)
IHC image of ab7254 staining in mouse alzheimer brain formalin fixed paraffin embedded tissue section, using MOM detection kit, ab127055. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab7254, 5µg/ml, for 15 mins at room temperature. DAB was used as the chromogen (ab103723). The section was counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ubiquitin antibody [Ubi-1] (ab7254)
Human normal placenta (ab29745). Staining is localised in the cytoplasm and in the nuclei. Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus, at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for mouse for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
Western blot - Anti-Ubiquitin antibody [Ubi-1] (ab7254)Image courtesy of an anonymous Abreview.
HeLa cells were co-transfected with a plasmid expressing a target protein together with Ubi expressing vector for 24 hours and either left untreated (Contr) or were treated with 10 µM MG-132 for 6 hours (+MG132). Then the protein of interest was pulled down using Flag agarose beads and and probed with ab7254 at a 1/2000 dilution. The secondary used was an Alexa-Fluor 680 conjugated goat anti-mouse polyclonal used at a 1/10000 dilution. The protein is known to be degraded through proteasome.