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Anti-YBX1 Antibody EP2708Y
Also for YBX1 (NM_004559)
|A synthetic peptide corresponding to residues near the C-terminus of human YB-1 was used as an immunogen.|
|Mouse, Rat, Human
||Lot dependent; please refer to CoA along with shipment
|WB, IHC, IF, IP, FC
||ICC/IF: Use a concentration of 5 ug/ml; WB: 1:10000 - 1:200000; IP: Use at an assay dependent concentration; IHC-P: 1:100 - 1:250; FC: 1:50 - 1:100
|PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%|
|Tissue culture supernatant
|Homo sapiens Y box binding protein 1 (YBX1)|
|BP-8; CSDA2; CSDB; DBPB; MDR-NF1; NSEP-1; NSEP1; YB-1; YB1|
Entrez Gene 4904 Human
Entrez Gene 22608 Mouse
Entrez Gene 500538 Rat
|Y-box-binding protein 1 (YBOX1, YBX1, YB-1) belongs to highly conserved Y-box protein family, a group of multifunctional gene regulators with cold shock domain (1). Targeting single stranded DNA and RNA, YB-1 regulates both translation and transcription in various genes. The expression of HLA class II genes is regulated by a series of cis-acting elements and trans-acting factors. As a cis-acting element, the Y-Box contains an inverted CCAAT box. An inverse correlation of YB-1 and HLA-DR beta chain mRNA levels suggests that YB-1 is a negative regulatory factor (2). Over-expression of YB-1 resulted in increased levels of PTP1B. Furthermore, depletion of YB-1 protein, by expression of a specific antisense construct, led to an approximately 70% decrease in expression of PTP1B, but no change in the level of its closest relative, TC-PTP. Data reveal an important role for YB-1 as a regulator of PTP1B expression, and further highlight PTP1B as a critical regulator of insulin- and cytokine-mediated signal transduction (3). Also, YB-1 has been implicated to activation of genes linked to cancer (e.g. cyclin A/B1, MMP-2, and MDR1) (4). However, it is shown to repress apoptosis related genes (e.g. p53 tumor suppressor) (5). YB-1 promotes separation of DNA strands that contains mismatches or is modified by cisplatin. Thus, YB-1 has been linked to possible role in DNA repair.|
|ES Cell Differentiation/IPSTranscription Factors |
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Western blot - YB1 antibody [EP2708Y]; All lanes : Anti-YB1 antibody [EP2708Y] at 1/200000 dilution.Lane 1 : HeLa cell lysate.Lane 2 : SW480 cell lysate.Lane 3 : A549 cell lysate.Lane 4 : MCF7 cell lysate.Lysates/proteins at 10 ug per lane.Secondary.goat anti-rabbit HRP at 1/1000 dilution.Predicted band size : 36 kDa.Observed band size : 50 kDa .
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - YB1 antibody [EP2708Y]; TA307283 at 1/100 dilution staining YB1 in human kidney by Immunohistochemistry, Paraffin-embedded tissue.
Immunocytochemistry/ Immunofluorescence-YB1 antibody [EP2708Y](TA307283); ICC/IF image of TA307283 stained HeLa cells. The cells were 4% formaldehyde 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 overnight at +4°C. The secondary antibody (green) was , DyLight 488 goat anti-rabbit IgG (H+L) 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 1.43uM.
Immunoprecipitation - Anti-YB1 antibody [EP2708Y]; YB1 was immunoprecipitated using 0.5mg Hek293 whole cell extract, 10ug of Rabbit monoclonal to YB1 and 50ul of protein G magnetic beads (+). No antibody was added to the control (-). .The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40ul SDS loading buffer and incubated for 10min at 70oC; 10ul of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with TA307283.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) .Band: 46kDa: YB1.
Flow Cytometry-YB1 antibody [EP2708Y](TA307283); Overlay histogram showing HeLa cells stained with TA307283 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody for 30 min at 22°C. The secondary antibody used was DyLight 488 goat anti-rabbit IgG (H+L) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1ug/1x10^6 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with methanol (5 min)/permeabilized with 0.1% PBS-Tween 20 used under the same conditions.