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Products:Tags & Cell Markers >> Cell Type Markers >> Tumor Associated
Overview
Product name
Anti-YB1 antibody
See all YB1 products (6) ...
Description
Rabbit polyclonal to YB1
Tested applications
ICC/IF, IHC-P, WB, IPmore details
Cross reactivity
Reacts with
Mouse, Rat, Human
Predicted to work with
Xenopus laevis
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human YB1.
(Peptide available as ab12411).
Positive control
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.
General notes
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.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Developmental Biology >> Embryogenesis >> Embryonic stem cells >> Surface molecules
Epigenetics and Nuclear Signaling >> Chromatin Binding Proteins >> DNA / RNA binding
Stem Cells >> Embryonic Stem Cells >> Intracellular
Epigenetics and Nuclear Signaling >> Transcription >> Other factors
Tags & Cell Markers >> Cell Type Markers >> Tumor Associated
Applications
Show applications key
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.
ICC/IF: Use a concentration of 1 µg/ml
IHC-P: 1/5000Perform 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 dilution.
Target
Function
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.
Sequence similarities
Contains 1 CSD (cold-shock) domain.
Post-translational
modifications
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.
Cellular localization
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.
Target information above from: UniProt accessionP67809
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- BP 8 antibody
- CBF-A antibody
- CCAAT binding transcription factor I subunit A antibody
see all
Database links
- Entrez Gene: 4904 Human
- Entrez Gene: 22608 Mouse
- Entrez Gene: 29206 Rat
- Entrez Gene: 500538 Rat
- Omim: 154030 Human
- SwissProt: P67809 Human
- SwissProt: P62960 Mouse
- SwissProt: Q810K5 Mouse
see all
Anti-YB1 antibody images:
Western blot - YB1 antibody (ab12148)
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
Western blot
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 kDa
Additional bands at : 50 kDa. We are unsure as to the identity of these extra bands.
A band of approx 50 KDa was partially blocked in several cell lines using a YB1 blocking peptide (ab12411). This suggests the antibody recognises YB1. Another band of approx 36 kDa also appears to be recognised, which may be a cleavage product of YB1.
Immunocytochemistry/ Immunofluorescence - YB1 antibody (ab12148)
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).
Immunocytochemistry/ Immunofluorescence - YB1 antibody (ab12148)
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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - YB1 antibody (ab12148)
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
Immunoprecipitation - YB1 antibody (ab12148)
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.
This image is courtesy of an anonymous Abreview
Immunocytochemistry/ Immunofluorescence - YB1 antibody (ab12148)
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.
References for Anti-YB1 antibody (ab12148)
This product has been referenced in:
- Narayan Vet al. A multiprotein binding interface in an intrinsically disordered region of the tumor suppressor protein interferon regulatory factor-1. J Biol Chem 286:14291-303 (2011).Read more (PubMed: 21245151) »
- Tsai BP, Wang X, Huang L, Waterman ML. Quantitative Profiling of In Vivo-assembled RNA-Protein Complexes Using a Novel Integrated Proteomic Approach Molecular & Cellular Proteomics 10:M110.007358 (2011).Read more (PubMed: ) »
See all 13 publications for this product
Publishing research using ab12148? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"