製品の概要

  • 製品名Anti-YB1 antibody
    YB1 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to YB1
  • アプリケーション適用あり: ICC/IF, IHC-P, WB, IPmore details
  • 種交差性
    交差種: Mouse, Rat, Human
    交差が予測される動物種: Xenopus laevis
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human YB1.

    (Peptide available as ab12411).

  • ポジティブ・コントロール
    • ab12148 gave a positive result in the following lysates: Hela, Jurkat, MCF-7, HEK293, NIH-3T3, MEF-1, PC12 Whole Cell Lysates; Hela Nuclear Lysate and HEK293 Whole Cell Lysate Transiently Overexpressing YB1.
  • 特記事項YB1 has a predicted band size of 36kDa. According to Evdolimova (1995) YB1 migrates by SDS-PAGE at 50kDa, which may be due to post-translational modification. YB1 is primarily detectable in the cytoplasm without any clear signal in nucleoli.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab12148 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 1 µg/ml.
IHC-P 1/5000. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
WB Use a concentration of 1 - 1.4 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 36 kDa). The 50 kDa band detected is consistent with the literature describing migration of YB1.
IP Use at an assay dependent concentration.

ターゲット情報

  • 機能Mediates pre-mRNA alternative splicing regulation. Binds to splice sites in pre-mRNA and regulates splice site selection. Binds and stabilizes cytoplasmic mRNA. Contributes to the regulation of translation by modulating the interaction between the mRNA and eukaryotic initiation factors (By similarity). Regulates the transcription of numerous genes. Its transcriptional activity on the multidrug resistance gene MDR1 is enhanced in presence of the APEX1 acetylated form at 'Lys-6' and 'Lys-7'. Binds to promoters that contain a Y-box (5'-CTGATTGGCCAA-3'), such as MDR1 and HLA class II genes. Promotes separation of DNA strands that contain mismatches or are modified by cisplatin. Has endonucleolytic activity and can introduce nicks or breaks into double-stranded DNA (in vitro). May play a role in DNA repair. Component of the CRD-mediated complex that promotes MYC mRNA stability.
    The secreted form acts as an extracellular mitogen and stimulates cell migration and proliferation.
  • 配列類似性Contains 1 CSD (cold-shock) domain.
  • 翻訳後修飾Ubiquitinated by RBBP6; leading to a decrease of YBX1 transcativational ability.
    In the absence of phosphorylation the protein is retained in the cytoplasm.
    Cleaved by a 20S proteasomal protease in response to agents that damage DNA. Cleavage takes place in the absence of ubiquitination and ATP. The resulting N-terminal fragment accumulates in the nucleus.
  • 細胞内局在Cytoplasm. Nucleus. Cytoplasmic granule. Secreted. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Shuttles between nucleus and cytoplasm. Predominantly cytoplasmic in proliferating cells. Cytotoxic stress and DNA damage enhance translocation to the nucleus. Localized with DDX1, MBNL1 and TIAL1 in stress granules upon stress. Secreted by mesangial and monocytic cells after inflammatory challenges. Translocates from the cytoplasm to the nucleus after and colocalizes with APEX1 in nuclear speckles after genotoxic stress.
  • Information by UniProt
  • 参照データベース
  • 別名
    • BP 8 antibody
    • CBF-A antibody
    • CCAAT binding transcription factor I subunit A antibody
    • CCAAT-binding transcription factor I subunit A antibody
    • CSDA2 antibody
    • CSDB antibody
    • DBPB antibody
    • DNA binding protein B antibody
    • DNA-binding protein B antibody
    • EFI-A antibody
    • Enhancer factor I subunit A antibody
    • MDR NF1 antibody
    • MGC104858 antibody
    • MGC110976 antibody
    • MGC117250 antibody
    • NSEP 1 antibody
    • NSEP1 antibody
    • Nuclease sensitive element binding protein 1 antibody
    • Nuclease-sensitive element-binding protein 1 antibody
    • p50 antibody
    • Q15905 antibody
    • Y-box binding protein 1 antibody
    • Y-box transcription factor antibody
    • Y-box-binding protein 1 antibody
    • YB 1 antibody
    • YB-1 antibody
    • YBOX1_HUMAN antibody
    • YBX 1 antibody
    • ybx1 antibody
    see all

Anti-YB1 antibody 画像

  • Anti-YB1 antibody (ab12148) at 1 µg/ml + HEK293 Whole Cell Lysate Transiently Overexpressing YB1 at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 36 kDa
    Observed band size : 50 kDa (why is the actual band size different from the predicted?)
    YB1 has a predicted band size of 36kDa based on its primary sequence (SwissProt). According to Evdolimova (1995) YB1 migrates by SDS-PAGE at 50kDa, which may be due to post-translational modification


  • Predicted band size : 36 kDa

    Lanes 1 to 10: Ab12148 - Rabbit polyclonal to YB1 (1.4µg/ml)     

    Lanes 1 to 10: 20µg of cell lysate per lane
    Lanes 6 to 10: blocking peptide used at 1µg/ml

    Lane 1: HeLa Nuclear Extract
    Lane 2: HeLa Whole Cell Lysate
    Lane 3: MCF-7 Cell Lysate
    Lane 4: Jurkat Whole Cell Lysate
    Lane 5: HEK293 Whole Cell Lysate
    Lane 6: HeLa Nuclear Extract + YB1 blocking peptide (ab12411)
    Lane 7: HeLa Whole Cell Lysate +  YB1 blocking peptide (ab12411)
    Lane 8: MCF-7 Cell Lysate +  YB1 blocking peptide (ab12411)
    Lane 9: Jurkat Whole Cell Lysate +  YB1 blocking peptide (ab12411)
    Lane 10: HEK293 Whole Cell Lysate +  YB1 blocking peptide (ab12411)

    Expected MW: 36 kDa

    A band of approx 36 kDa was partially blocked in several cell lines using a YB1 blo

  • ICC/IF image of ab12148 stained human HeLa cells. The cells were PFA fixed (3.7% PFA, 5 min) and incubated with the antibody (ab12148, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • ICC/IF image of ab12148 stained human HeLa cells. The cells were PFA fixed (3.7% PFA, 5 min) and incubated with the antibody (ab12148, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • Image courtesy of Human Protein Atlas

    ab12148 staining YB1 in Human duodenum, showing a distinct and strong staining pattern of the glandular cells. Paraffin embedded human duodenum was incubated with ab12148 (1/5000 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

    ab12148 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org

  • ab12148 was diluted 1/500 and incubated with A2780 whole cell lysate and protein A/G matrix for 16 hours at 4°C to achieve immunoprecipitation. 200 µg of protein was present in the lysate input.  Lane 2 shows results of a different antibody used as the  negative control.  An HRP-conjugated goat anti-rabbit antibody was used for the western blot step.

    See Abreview

  • ICC/IF image of ab12148 stained Hela cells. The cells were 4% PFA fixed (10 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 (ab12148, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 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 1.43µM.

Anti-YB1 antibody (ab12148) 使用論文

This product has been referenced in:
  • ErLin S  et al. Musashi-1 maintains blood-testis barrier structure during spermatogenesis and regulates stress granule formation upon heat stress. Mol Biol Cell 26:1947-56 (2015). Mouse . Read more (PubMed: 25717188) »
  • Jefferson M  et al. Host factors that interact with the pestivirus N-terminal protease, Npro, are components of the ribonucleoprotein complex. J Virol 88:10340-53 (2014). ICC/IF . Read more (PubMed: 24965446) »

See all 22 Publications for this product

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Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (293T)
Loading amount 30000 cells
Specification 293T
Gel Running Conditions Reduced Denaturing (4-20% Tris-Glycine)
Blocking step 5% Milk, 1% BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

投稿 Feb 26 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Human Renal Proximal Tubular cells)
Loading amount 10 µg
Specification Human Renal Proximal Tubular cells
Gel Running Conditions Reduced Denaturing (7.5 %)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Dr. Robert Jenkins

Verified customer

投稿 Apr 30 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Renal proximal tubular epithelial cells)
Specification Renal proximal tubular epithelial cells
Fixative Paraformaldehyde
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Dr. Robert Jenkins

Verified customer

投稿 Mar 26 2009

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Tissue lysate - whole (Mouse heart tissue)
Loading amount 20 µg
Specification Mouse heart tissue
Gel Running Conditions Reduced Denaturing (10% gel)
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 Nov 12 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Mouse Tissue lysate - whole (Mouse heart tissue)
Total protein in input 200 µg
Specification Mouse heart tissue
Immuno-precipitation step Protein A/G
Username

Abcam user community

Verified customer

投稿 Nov 12 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Mouse Tissue lysate - whole (lung tissue)
Total protein in input 300 µg
Specification lung tissue
Immuno-precipitation step Protein A/G
Username

Abcam user community

Verified customer

投稿 Sep 23 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Rat Tissue lysate - whole (lung tissue)
Total protein in input 300 µg
Specification lung tissue
Immuno-precipitation step Protein A/G
Username

Abcam user community

Verified customer

投稿 Sep 23 2008

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