概述

  • 产品名称
  • 描述
    兔多克隆抗体to YB1
  • 经测试应用
    适用于: ICC/IF, IHC-P, WB, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: Xenopus laevis
  • 免疫原

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

    (Peptide available as ab12411).

  • 阳性对照
    • WB: HeLa, Jurkat, MCF-7, HEK293, NIH-3T3, MEF-1 and PC12 whole cell lysates, HeLa nuclear lysate and HEK293 whole cell lysate transiently overexpressing YB1. ICC/IF: HeLa cells. IHC-P: Human duodenum tissue.
  • 常规说明
    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.

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能
    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.
  • 序列相似性
    Contains 1 CSD (cold-shock) domain.
  • 翻译后修饰
    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.
  • 细胞定位
    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
  • 数据库链接
  • 别名
    • 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

图片

  • All lanes : Anti-YB1 antibody (ab12148) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 3 : Jurkat (Human) Whole Cell Lysate
    Lane 4 : T47D whole cell lysate (ab14899)
    Lane 5 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 36 kDa
    Observed band size : 50 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 100 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab12148 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • 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-iT™ 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.

  • 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
  • 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-iT™ 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).

  • All lanes : Anti-YB1 antibody (ab12148) at 1.4 µg/ml

    Lane 1 : HeLa Nuclear lysate
    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 lysate with YB1 peptide (ab12411) at 1 µg/ml
    Lane 7 : HeLa Whole cell lysate with YB1 peptide (ab12411) at 1 µg/ml
    Lane 8 : MCF-7 cell lysate with YB1 peptide (ab12411) at 1 µg/ml
    Lane 9 : Jurkat whole cell lysate with YB1 peptide (ab12411) at 1 µg/ml
    Lane 10 : HEK293 whole cell lysate with YB1 peptide (ab12411) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.


    Predicted band size : 36 kDa
    Observed band size : 36,50 kDa (why is the actual band size different from the predicted?)
  • 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.

  • 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

文献

This product has been referenced in:
  • Eliscovich C  et al. Imaging mRNA and protein interactions within neurons. Proc Natl Acad Sci U S A 114:E1875-E1884 (2017). WB, ICC/IF . Read more (PubMed: 28223507) »
  • Malchenko S  et al. Stabilization of HIF-1a and HIF-2a, up-regulation of MYCC and accumulation of stabilized p53 constitute hallmarks of CNS-PNET animal model. PLoS One 12:e0173106 (2017). IHC-P ; Human . Read more (PubMed: 28249000) »

See all 38 Publications for this product

客户评价及客户问答

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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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 Sep 23 2008

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