Anti-YB1 antibody (ab12148)
Key features and details
- Rabbit polyclonal to YB1
- Suitable for: ICC/IF, WB
- Reacts with: Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
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- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-YB1 antibody
See all YB1 primary antibodies -
Description
Rabbit polyclonal to YB1 -
Host species
Rabbit -
Specificity
Replenishment batches of our polyclonal antibody, ab12148 are tested in WB. Previous batches were additionally validated in ICC/IF. This application is still expected to work and is covered by our Abpromise guarantee. You may also be interested in our alternative recombinant antibody, ab76149.
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Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Xenopus laevis -
Immunogen
Synthetic peptide corresponding to Human YB1 aa 1-100 conjugated to keyhole limpet haemocyanin.
(Peptide available asab12411) -
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.The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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ICC/IF | (3) |
Use a concentration of 1 µg/ml.
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WB | (9) |
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.
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Notes |
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ICC/IF
Use a concentration of 1 µg/ml. |
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. |
Target
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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
modificationsUbiquitinated 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. - Information by UniProt
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Database links
- Entrez Gene: 4904 Human
- Entrez Gene: 22608 Mouse
- Entrez Gene: 500538 Rat
- Omim: 154030 Human
- SwissProt: P67809 Human
- SwissProt: P62960 Mouse
- SwissProt: P62961 Rat
- SwissProt: P21573 Xenopus laevis
see all -
Alternative names
- BP 8 antibody
- CBF-A antibody
- CCAAT binding transcription factor I subunit A antibody
see all
Images
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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 :T-47D 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
All lanes : 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 minutesThis 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.
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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).
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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).
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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 withYB1 peptide (ab12411) at 1 µg/ml
Lane 7 : HeLa Whole cell lysate withYB1 peptide (ab12411) at 1 µg/ml
Lane 8 : MCF-7 cell lysate withYB1 peptide (ab12411) at 1 µg/ml
Lane 9 : Jurkat whole cell lysate withYB1 peptide (ab12411) at 1 µg/ml
Lane 10 : HEK293 whole cell lysate withYB1 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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (87)
ab12148 has been referenced in 87 publications.
- Feng X et al. Long noncoding RNA Gm31629 protects against mucosal damage in experimental colitis via YB-1/E2F pathway. JCI Insight 7:N/A (2022). PubMed: 35143419
- Gao W et al. SIAH1 reverses chemoresistance in epithelial ovarian cancer via ubiquitination of YBX-1. Oncogenesis 11:13 (2022). PubMed: 35273154
- Hatstat AK et al. Characterization of Small-Molecule-Induced Changes in Parkinson's-Related Trafficking via the Nedd4 Ubiquitin Signaling Cascade. Cell Chem Biol 28:14-25.e9 (2021). PubMed: 33176158
- Taylor L et al. Y-Box Binding Protein-1: A Neglected Target in Pediatric Brain Tumors? Mol Cancer Res 19:375-387 (2021). PubMed: 33239357
- Mateu-Regué À et al. Unveiling mRNP composition by fluorescence correlation and cross-correlation spectroscopy using cell lysates. Nucleic Acids Res 49:e119 (2021). PubMed: 34478550