製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to Ubiquitin
  • アプリケーション
    適用あり: ICC/IF, IHC-FoFr, IHC-P, WB, IPmore details
  • 種交差性
    交差種: Mouse, Rat, Human, African green monkey
    交差が予測される動物種: Horse, Cow
  • 免疫原

    Recombinant full length protein corresponding to Human Ubiquitin.

  • ポジティブ・コントロール
    • Pancreas and Alzheimer’s brain Rat brain
  • 特記事項

    This antibody stains the periphery of senile plaques and of neurofibrillary tangles in Alzheimer's disease brain, the Lewy bodies in Parkinson's disease brain, and the Mallory bodies in alcoholic liver disease.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab7780 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 at an assay dependent concentration.

1/500 - 1/1000.

IHC-FoFr 1/50 - 1/100.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Boil tissue sections in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min.
WB Use at an assay dependent concentration. Predicted molecular weight: 8.5 kDa. Used at a dilution of 1/2000 B16F10 murine melanoma cell line (see Abreview). Predicted molecular weight: 8.5 kDa.
IP Use at an assay dependent concentration.

ターゲット情報

  • 関連性
    Function: Ubiquitin exists either covalently attached to another protein, or free (unanchored). When covalently bound, it is conjugated to target proteins via an isopeptide bond either as a monomer (monoubiquitin), a polymer linked via different Lys residues of the ubiquitin (polyubiquitin chains) or a linear polymer linked via the initiator Met of the ubiquitin (linear polyubiquitin chains). Polyubiquitin chains, when attached to a target protein, have different functions depending on the Lys residue of the ubiquitin that is linked: Lys-6-linked may be involved in DNA repair; Lys-11-linked is involved in ERAD (endoplasmic reticulum-associated degradation) and in cell-cycle regulation; Lys-29-linked is involved in lysosomal degradation; Lys-33-linked is involved in kinase modification; Lys-48-linked is involved in protein degradation via the proteasome; Lys-63-linked is involved in endocytosis, DNA-damage responses as well as in signaling processes leading to activation of the transcription factor NF-kappa-B. Linear polymer chains formed via attachment by the initiator Met lead to cell signaling. Ubiquitin is usually conjugated to Lys residues of target proteins, however, in rare cases, conjugation to Cys or Ser residues has been observed. When polyubiquitin is free (unanchored-polyubiquitin), it also has distinct roles, such as in activation of protein kinases, and in signaling. Similarity: Belongs to the ubiquitin family. Contains 3 ubiquitin-like domains.
  • 細胞内局在
    Cell Membrane, Cytoplasmic and Nuclear
  • 参照データベース
  • 別名
    • Epididymis secretory protein Li 50 antibody
    • FLJ25987 antibody
    • HEL S 50 antibody
    • MGC8385 antibody
    • Polyubiquitin B antibody
    • RPS 27A antibody
    • RPS27A antibody
    • UBA 52 antibody
    • UBA 80 antibody
    • UBA52 antibody
    • UBA80 antibody
    • UBB antibody
    • UBB_HUMAN antibody
    • UBC antibody
    • UBCEP 1 antibody
    • UBCEP 2 antibody
    • UBCEP1 antibody
    • UBCEP2 antibody
    • Ubiquitin antibody
    • Ubiquitin B antibody
    see all

画像


  • Developed using the ECL technique

    Predicted band size : 8.5 kDa


    Exposure time : 30 seconds

    Image courtesy of an anonymous Abreview.

    ab7780 used in Western Blot. Whole cell lysate prepared from human lung cancer cell lines was loaded at 100000 cells. The gel was run under reduced, denaturing conditions. 5% milk used as the blocking agent. ab7780 used at a 1/1000 dilution, incubated for 16 hours at 4°C in 5% BSA in TBS-Tween (0.1%). The secondary was an HRP conjugated mouse anti-rabbit, used at a 1/10,000 dilution.Specific bands seen : 16, 24, 32, 50, 60, kDa.

  • All lanes : Anti-Ubiquitin antibody (ab7780) at 1/2000 dilution

    Lane 1 : Human Panc-1 whole cell lysate
    Lane 2 : Human Panc-1 whole cell lysate

    Lysates/proteins at 700 µg per lane.

    Secondary
    HRP Donkey anti-rabbit IgG polyclonal at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 8.5 kDa


    Exposure time : 25 seconds

    This image is courtesy of an Abreview submitted by Armen Petrosyan

    Ubiquitin western blot of the complexes pulled down with anti-c-Myc antibodies from the lysates of cells treated with DMSO or MG-132. Treatment with the proteasome inhibitor, MG-132, increases ubiquitinated protein; seen as several bands between 55-72 kDa.

    See Abreview

  • ab7780 staining Ubiquitin in Panc-1 cells by Immunocytochemistry/ Immunofluorescence.Cells were fixed in formaldehyde, blocked with 1% serum for 1 hour at 22°C and then incubated with ab7780 at a 1/2000 dilution for 1 hour at 22°C. The secondary was used at a 1/200 dilution.Green - Golgi residental proteins C2GnT-M.Red -Ubiquitin.Blue - nucleus staining by DAPI.

    See Abreview

  • ab7780 staining ubiquitin in cells transfected with FGAMS-EGFP by ICC/IF (immunocytochemistry/immunofluorescence). Cells were fixed with 3.7% methanol-free formaldehyde at 37°C for 15-20 minutes, blocked with 5% goat serum in PBS-T buffer for 30-60 minutes at room temperature. An Alexa Fluor® 594-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody

  • ICC/IF image of ab7780 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum (ab7481) / 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 (ab7780, 1æg/ml) overnight at +4øC. The secondary antibody (green)ÿwas Alexa Fluor© 488 goat anti-mouse IgG (H+L) ab150113) used at a 1/1000 dilution for 1h. Alexa Fluor© 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43æM.

  • ab7780 (1/100) staining Ubiquitin in paraffin-embedded brain sections of a patient with Alzheimer's disease.

  • ab7780 staining ubiquitin in Mouse kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 1 hour at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/90) for 16 hours at 4°C. A HRP-conjugated Goat anti-rabbit polyclonal (1/1000) was used as the secondary antibody.

    See Abreview

  • ab7780 at a dilution of 1/1000, staining Ubiquitin in neurons (Alexa 488 secondary at 1/2000) on 30µm coronal rat brain (ab29475) tissue sections in free floating IHC (see protocol link for detailed description). Image shows neuronal staining observed with [A] 20x objective and [B] 40x objective.

    NB: No labeling observed following omission of primary antibody.

    Sections were viewed using an Axioplan 2 Imaging microscope (Imaging Associates) fitted with 10x, 20x and 40x Plan-Neofluorobjectives (Zeiss, Germany) and images were taken using a AxioCam Hrm digital camera (Zeiss, Germany) and AxioVision software (Imaging Associates).

参考文献

This product has been referenced in:
  • Ye M  et al. Curcumin Improves Palmitate-Induced Insulin Resistance in Human Umbilical Vein Endothelial Cells by Maintaining Proteostasis in Endoplasmic Reticulum. Front Pharmacol 8:148 (2017). Human . Read more (PubMed: 28377722) »
  • Bobo-Jiménez V  et al. APC/C(Cdh1)-Rock2 pathway controls dendritic integrity and memory. Proc Natl Acad Sci U S A 114:4513-4518 (2017). WB ; Mouse . Read more (PubMed: 28396402) »

See all 60 Publications for this product

レビューと Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Tissue (Vascular smooth muscle cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Treatment
100 uM H2O2 for 2 hours
Specification
Vascular smooth muscle cells
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 May 22 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Rat hepatocytes)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
Rat hepatocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Username

Dr. Armen Petrosyan

Verified customer

投稿 Apr 03 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (Human mast cell)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
10 µg
Specification
Human mast cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Seong Hoon Kim

Verified customer

投稿 Jun 23 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - other (Mitochondrial fraction from skeletal muscle)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Mitochondrial fraction from skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Feb 29 2016

Application
Western blot
Sample
Mouse Tissue lysate - other (Mitochondrial fraction from skeletal muscle)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
20 µg
Specification
Mitochondrial fraction from skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Feb 18 2016

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Human Cell lysate - whole cell (T lymphocyte)
Specification
T lymphocyte
Treatment
No chemical
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Dec 10 2014

Application
Western blot
Loading amount
1000 µg
Gel Running Conditions
Reduced Denaturing (10% gel)
Sample
Human Cell lysate - whole cell (Brain cancer cells (immunoprecipitated lysates))
Specification
Brain cancer cells (immunoprecipitated lysates)
Treatment
proteasome inhibitor MG-132
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

投稿 Nov 22 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Sample
Human Cell (HS 5)
Specification
HS 5
Permeabilization
Yes - Triton X 100
Fixative
Formaldehyde
Username

Mr. Kannan Govindaraj

Verified customer

投稿 Aug 02 2013



We do not have any experimental data regarding the cross-reactivity of ab7780 with SUMO proteins. I compared the immunogen for ab7780 with the sequences of SUMO 1, 2, 3, 4 (P63165, P61956, P55854, Q6EEV6-Uniprot) and the percentage homology i...

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I am happy to hear that the antibody is working for you when using BSA instead of milk! If you have any additional questions, please let me know and I will be happy to help you further.

1-10 of 46 Abreviews or Q&A

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

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