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RecombinantRabMAb

Recombinant Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)

Submit a review Q&A (1)References (21)
Western blot - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
  • Immunocytochemistry/ Immunofluorescence - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
  • Flow Cytometry (Intracellular) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
  • Western blot - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
  • Immunoprecipitation - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
  • Western blot - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
  • Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3103Y] to Cullin 1/CUL-1
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

APC Carrier Free HRP PE

Overview

  • Product name

    Anti-Cullin 1/CUL-1 antibody [EPR3103Y]
    See all Cullin 1/CUL-1 primary antibodies

  • Description

    Rabbit monoclonal [EPR3103Y] to Cullin 1/CUL-1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Cullin 1/CUL-1 aa 750 to the C-terminus (C terminal). The exact sequence is proprietary.

  • Positive control

    • HeLa, MCF7, A549 and PC12 cell lysates; human cervical carcinoma tissue.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab75817 in the following tested applications.

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

Application Abreviews Notes
Flow Cyt (Intra)
1/20.
ICC/IF
1/100 - 1/250.
WB
1/1000. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa).

For unpurified, use 1/2500 - 1/5000.
IP
1/10 - 1/50.
IHC-P
1/50 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Notes
Flow Cyt (Intra)
1/20.
ICC/IF
1/100 - 1/250.
WB
1/1000. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa).

For unpurified, use 1/2500 - 1/5000.
IP
1/10 - 1/50.
IHC-P
1/50 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • Function

    Core component of multiple cullin-RING-based SCF (SKP1-CUL1-F-box protein) E3 ubiquitin-protein ligase complexes, which mediate the ubiquitination of proteins involved in cell cycle progression, signal transduction and transcription. In the SCF complex, serves as a rigid scaffold that organizes the SKP1-F-box protein and RBX1 subunits. May contribute to catalysis through positioning of the substrate and the ubiquitin-conjugating enzyme. The E3 ubiquitin-protein ligase activity of the complex is dependent on the neddylation of the cullin subunit and is inhibited by the association of the deneddylated cullin subunit with TIP120A/CAND1. The functional specificity of the SCF complex depends on the F-box protein as substrate recognition component. SCF(BTRC) and SCF(FBXW11) direct ubiquitination of CTNNB1 and participate in Wnt signaling. SCF(FBXW11) directs ubiquitination of phosphorylated NFKBIA. SCF(BTRC) directs ubiquitination of NFKBIB, NFKBIE, ATF4, SMAD3, SMAD4, CDC25A, FBXO5 and probably NFKB2. SCF(SKP2) directs ubiquination of phosphorylated CDKN1B/p27kip and is involved in regulation of G1/S transition. SCF(SKP2) directs ubiquination of ORC1, CDT1, RBL2, ELF4, CDKN1A, RAG2, FOXO1A, and probably MYC and TAL1. SCF(FBXW7) directs ubiquitination of cyclin E, NOTCH1 released notch intracellular domain (NICD), and probably PSEN1. SCF(FBXW2) directs ubiquitination of GCM1. SCF(FBXO32) directs ubiquitination of MYOD1. SCF(FBXO7) directs ubiquitination of BIRC2 and DLGAP5. SCF(FBXO33) directs ubiquitination of YBX1. SCF(FBXO11) does not seem to direct ubiquitination of TP53. SCF(BTRC) mediates the ubiquitination of NFKBIA at 'Lys-21' and 'Lys-22'; the degradation frees the associated NFKB1-RELA dimer to translocate into the nucleus and to activate transcription. SCF(Cyclin F) directs ubiquitination of CP110.

  • Tissue specificity

    Expressed in lung fibroblasts.

  • Pathway

    Protein modification; protein ubiquitination.

  • Sequence similarities

    Belongs to the cullin family.

  • Post-translational
    modifications

    Neddylated; which enhances the ubiquitination activity of SCF. Deneddylated via its interaction with the COP9 signalosome (CSN) complex. Deneddylated by Epstein-Barr virus BPLF1 leading to a S-phase-like environment that is required for efficient replication of the viral genome.
  • Information by UniProt

  • Database links

    • Alternative names

      • CUL 1 antibody
      • CUL-1 antibody
      • CUL1 antibody
      • CUL1_HUMAN antibody
      • Cullin-1 antibody
      • Cullin1 antibody
      • MGC149834 antibody
      • MGC149835 antibody
      see all

    Images

    • Western blot - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      Western blot - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      All lanes : Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817) at 1/1000 dilution (purified)

      Lane 1 : HeLa cell lysate
      Lane 2 : MCF7 cell lysate
      Lane 3 : A549 cell lysate
      Lane 4 : MDA-MB-435 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

      Predicted band size: 90 kDa
      Observed band size: 90 kDa



      Blocking buffer: 5% NFDM/TBST
      Dilution buffer: 5% NFDM/TBST

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      Immunohistochemical staining of paraffin embedded human lung carcinoma with purified ab75817 at a working dilution of 1/50. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
    • Immunocytochemistry/ Immunofluorescence - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      Immunocytochemistry/ Immunofluorescence - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)

      Immunofluorescence staining of HeLa cells with purified ab75817 at a working dilution of 1/1000, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab75817 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

    • Flow Cytometry (Intracellular) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      Flow Cytometry (Intracellular) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)

      Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Cullin 1/CUL-1 with purified ab75817 at 1/20 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr®488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. 

       

       

    • Western blot - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      Western blot - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      All lanes : Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817) at 1/1000 dilution (purified)

      Lane 1 : PC-12 cell lysate
      Lane 2 : NIH/3T3 cell lysate
      Lane 3 : mouse brain cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

      Predicted band size: 90 kDa
      Observed band size: 90 kDa



      Blocking buffer: 5% NFDM/TBST
      Dilution buffer: 5% NFDM/TBST

    • Immunoprecipitation - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      Immunoprecipitation - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)

      ab75817 (purified) at 1/120 immunoprecipitating Cullin 1 in 10 μg MCF7 whole cell lysate (Lanes 1 and 2, observed at 90 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab75817 at a working dilution of 1/50. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)

      Unpurified ab75817 at 1/100 dilution staining Cullin 1/CUL-1 in human cervical carcinoma by Immunohistochemistry, Paraffin-embedded tissue.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Western blot - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      Western blot - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      All lanes : Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817) at 1/5000 dilution (unpurified)

      Lane 1 : HeLa cell lysate
      Lane 2 : MCF7 cell lysate
      Lane 3 : A549 cell lysate
      Lane 4 : PC12 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : goat anti-rabbit HRP at 1/2000 dilution

      Predicted band size: 90 kDa
      Observed band size: 90 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)

      Unpurified ab75817 showing positive staining in Normal human tonsil tissue.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)

      Unpurified ab75817 showing positive staining in human Urinary bladder transitional carcinoma tissue.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)

      Unpurified ab75817 showing positive staining in human Ovarian carcinoma tissue.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)
      Anti-Cullin 1/CUL-1 antibody [EPR3103Y] (ab75817)

    References (21)

    Publishing research using ab75817? Please let us know so that we can cite the reference in this datasheet.

    ab75817 has been referenced in 21 publications.

    • Zhang W  et al. Tumor-associated antigen Prame targets tumor suppressor p14/ARF for degradation as the  receptor protein of CRL2Prame complex. Cell Death Differ 28:1926-1940 (2021). PubMed: 33504946
    • Dong X  et al. Tumor suppressor DCAF15 inhibits epithelial-mesenchymal transition by targeting ZEB1 for proteasomal degradation in hepatocellular carcinoma. Aging (Albany NY) 13:10603-10618 (2021). PubMed: 33833131
    • Du MG  et al. The Absence of PTEN in Breast Cancer Is a Driver of MLN4924 Resistance. Front Cell Dev Biol 9:667435 (2021). PubMed: 33996822
    • Sweeney MA  et al. The ubiquitin ligase Cullin-1 associates with chromatin and regulates transcription of specific c-MYC target genes. Sci Rep 10:13942 (2020). PubMed: 32811853
    • Zhou L  et al. Induction of NEDD8-conjugating enzyme E2 UBE2F by platinum protects lung cancer cells from apoptosis and confers to platinum-insensitivity. Cell Death Dis 11:975 (2020). PubMed: 33184273
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Question

    My question is which peptides did you use to raise this antibody? Were they from the N-terminal? Many thanks,

    Read More
    Answer

    Thank you for your enquiry. The peptide used to raise this Cullin 1 antibody is from the C-terminal region, within the last 20 aa. Note that this is a general region of the immunogen sequence and the exact sequence information is proprietary. I hope this information will still be helpful. If there is anything else that I can help you with, please do not hesitate to contact me.

    Read More

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