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Anti-YB1 antibody [EP2708Y] ab76149 is a rabbit monoclonal antibody that is used in YB1 western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EP2708Y is the most widely used clone for YB1 on the market and is cited in >80 publications
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- One antibody for all your YB1 staining, use in YB1 western blotting, IHC, immunofluorescence and flow cytometry


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YB1 antibody [EP2708Y] (AB76149), expandable thumbnail
  • Western blot - Anti-YB1 antibody [EP2708Y] (AB76149), expandable thumbnail
  • Western blot - Anti-YB1 antibody [EP2708Y] (AB76149), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YB1 antibody [EP2708Y] (AB76149), expandable thumbnail
  • Western blot - Anti-YB1 antibody [EP2708Y] (AB76149), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Tested
Expected
Tested
Expected
Expected
Rat
Tested
Expected
Tested
Expected
Expected

Tested
Tested

Species

Mouse

Dilution info

1/50 - 1/250

Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Species

Rat

Dilution info

1/50 - 1/250

Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Species

Human

Dilution info

1/50 - 1/250

Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species

Human

Dilution info

1/30

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/1000

Notes

For unpurified use at 1/10000 - 1/20000.

Species

Rat

Dilution info

1/1000

Notes

For unpurified use at 1/10000 - 1/20000.

Species

Human

Dilution info

1/1000

Notes

For unpurified use at 1/10000 - 1/20000.

Tested
Tested

Species

Human

Dilution info

1/100

Notes

For unpurified use at 5μg/ml.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/50 - 1/100

Notes

Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

8 products for Alternative Product

Target data

Function

DNA- and RNA-binding protein involved in various processes, such as translational repression, RNA stabilization, mRNA splicing, DNA repair and transcription regulation (PubMed:8188694, PubMed:10817758, PubMed:11698476, PubMed:14718551, PubMed:18809583, PubMed:31358969). Predominantly acts as a RNA-binding protein: binds preferentially to the 5'-[CU]CUGCG-3' RNA motif and specifically recognizes mRNA transcripts modified by C5-methylcytosine (m5C) (PubMed:19561594, PubMed:31358969). Promotes mRNA stabilization: acts by binding to m5C-containing mRNAs and recruiting the mRNA stability maintainer ELAVL1, thereby preventing mRNA decay (PubMed:10817758, PubMed:11698476, PubMed:31358969). Component of the CRD-mediated complex that promotes MYC mRNA stability (PubMed:19029303). Contributes to the regulation of translation by modulating the interaction between the mRNA and eukaryotic initiation factors (By similarity). Plays a key role in RNA composition of extracellular exosomes by defining the sorting of small non-coding RNAs, such as tRNAs, Y RNAs, Vault RNAs and miRNAs (PubMed:27559612, PubMed:29073095). Probably sorts RNAs in exosomes by recognizing and binding C5-methylcytosine (m5C)-containing RNAs (PubMed:28341602, PubMed:29073095). Acts as a key effector of epidermal progenitors by preventing epidermal progenitor senescence: acts by regulating the translation of a senescence-associated subset of cytokine mRNAs, possibly by binding to m5C-containing mRNAs (PubMed:29712925). Also involved in pre-mRNA alternative splicing regulation: binds to splice sites in pre-mRNA and regulates splice site selection (PubMed:12604611). Also able to bind DNA: regulates transcription of the multidrug resistance gene MDR1 is enhanced in presence of the APEX1 acetylated form at 'Lys-6' and 'Lys-7' (PubMed:18809583). Binds to promoters that contain a Y-box (5'-CTGATTGGCCAA-3'), such as MDR1 and HLA class II genes (PubMed:8188694, PubMed:18809583). Promotes separation of DNA strands that contain mismatches or are modified by cisplatin (PubMed:14718551). Has endonucleolytic activity and can introduce nicks or breaks into double-stranded DNA, suggesting a role in DNA repair (PubMed:14718551). The secreted form acts as an extracellular mitogen and stimulates cell migration and proliferation (PubMed:19483673).

Alternative names

Y-box-binding protein 1, YB-1, CCAAT-binding transcription factor I subunit A, DNA-binding protein B, Enhancer factor I subunit A, Nuclease-sensitive element-binding protein 1, Y-box transcription factor, CBF-A, DBPB, EFI-A, NSEP1, YBX1, YB1

Recommended products

Anti-YB1 antibody [EP2708Y] ab76149 is a rabbit monoclonal antibody that is used in YB1 western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EP2708Y is the most widely used clone for YB1 on the market and is cited in >80 publications
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- One antibody for all your YB1 staining, use in YB1 western blotting, IHC, immunofluorescence and flow cytometry

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EP2708Y

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Y-box Binding Protein 1 (YB1) also known as YBX1 is a multifunctional protein with a mass of approximately 36 kDa. It belongs to the cold shock protein family that contains a highly conserved cold shock domain. YB1 is broadly expressed across many tissues predominantly found in the cytoplasm and nucleus where it functions as a transcription factor and RNA-binding protein. Through its transcriptional regulatory activity YB1 can bind to Y-box sequences in the promoter regions of genes and modulate their expression.

Biological function summary

YB1 plays vital roles in regulating cell proliferation stress response and differentiation. It does not operate alone; it interacts with various complexes notably ribonucleoprotein complexes. YB1 regulates gene expression at both transcriptional and translational levels impacting cell cycle progression and apoptosis pathways. Its ability to bind RNA and DNA makes it integral in controlling the expression of genes related to stress responses and developmental processes.

Pathways

YB1 is essential in many signaling pathways that govern cell growth and survival including the PI3K/AKT pathway and MAPK pathway. In these pathways it interacts with proteins like AKT1 and MAPK3 influencing cellular responses to external and internal stimuli. These interactions reveal the adaptability of YB1 in various cellular contexts allowing it to mediate pathway-specific responses to environmental cues thereby promoting the maintenance of cellular homeostasis and adaptation.

Associated diseases and disorders

The dysregulation of YB1 has been linked to cancer and neurodegenerative diseases. Overexpression or altered localization of YB1 often in connection with other proteins such as p53 contributes to tumorigenesis by enhancing cell proliferation and inhibiting programmed cell death. In neurodegenerative diseases impaired YB1 function can negatively affect neuronal survival potentially mediated through its interactions with proteins like tau which are implicated in conditions such as Alzheimer's disease. Understanding of these associations highlights the potential of YB1 as a therapeutic target in disease management.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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14 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YB1 antibody [EP2708Y] (ab76149), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YB1 antibody [EP2708Y] (ab76149)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat stomach tissue labelling YB1 with purified ab76149 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Western blot - Anti-YB1 antibody [EP2708Y] (ab76149), expandable thumbnail

    Western blot - Anti-YB1 antibody [EP2708Y] (ab76149)

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-YB1 antibody [EP2708Y] (ab76149) at 1/1000 dilution

    Lane 1: NIH/3T3 whole cell lysate at 10 µg

    Lane 2: Raw264.7 whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

    Predicted band size: 36 kDa

    Observed band size: 50 kDa

  • Western blot - Anti-YB1 antibody [EP2708Y] (ab76149), expandable thumbnail

    Western blot - Anti-YB1 antibody [EP2708Y] (ab76149)

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-YB1 antibody [EP2708Y] (ab76149) at 1/10000 dilution

    Lane 1: C6 whole cell lysate at 10 µg

    Lane 2: PC-12 whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

    Predicted band size: 36 kDa

    Observed band size: 50 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YB1 antibody [EP2708Y] (ab76149), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YB1 antibody [EP2708Y] (ab76149)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue labelling YB1 with purified ab76149 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Western blot - Anti-YB1 antibody [EP2708Y] (ab76149), expandable thumbnail

    Western blot - Anti-YB1 antibody [EP2708Y] (ab76149)

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-YB1 antibody [EP2708Y] (ab76149) at 1/1000 dilution

    Lane 1: HeLa whole cell lysate at 10 µg

    Lane 2: SW480 whole cell lysate at 10 µg

    Lane 3: A549 whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution

    Predicted band size: 36 kDa

    Observed band size: 50 kDa

  • Immunoprecipitation - Anti-YB1 antibody [EP2708Y] (ab76149), expandable thumbnail

    Immunoprecipitation - Anti-YB1 antibody [EP2708Y] (ab76149)

    ab76149 (purified) at 1/30 immunoprecipitating YB1 in MCF-7 whole cell lysate.

    Lane 1 (input): MCF-7 whole cell lysate (10μg)

    Lane 2 (+): ab76149 + MCF-7 whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab76149 in MCF-7 whole cell lysate.

    For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    All lanes: Immunoprecipitation - Anti-YB1 antibody [EP2708Y] (ab76149)

    Predicted band size: 36 kDa

    Observed band size: 50 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YB1 antibody [EP2708Y] (ab76149), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YB1 antibody [EP2708Y] (ab76149)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labelling YB1 with purified ab76149 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunoprecipitation - Anti-YB1 antibody [EP2708Y] (ab76149), expandable thumbnail

    Immunoprecipitation - Anti-YB1 antibody [EP2708Y] (ab76149)

    ab76149 (purified) at 1/30 immunoprecipitating YB1 in HeLa whole cell lysate.

    Lane 1 (input): HeLa whole cell lysate (10μg)

    Lane 2 (+): ab76149 + HeLa whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab76149 in HeLa whole cell lysate.

    For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    All lanes: Immunoprecipitation - Anti-YB1 antibody [EP2708Y] (ab76149)

    Predicted band size: 36 kDa

    Observed band size: 50 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-YB1 antibody [EP2708Y] (ab76149), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-YB1 antibody [EP2708Y] (ab76149)

    Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling YB1 with purified ab76149 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

  • Flow Cytometry (Intracellular) - Anti-YB1 antibody [EP2708Y] (ab76149), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-YB1 antibody [EP2708Y] (ab76149)

    Intracellular Flow Cytometry analysis of HeLa cells labelling YB1 with purified ab76149 at 1/90 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Immunoprecipitation - Anti-YB1 antibody [EP2708Y] (ab76149), expandable thumbnail

    Immunoprecipitation - Anti-YB1 antibody [EP2708Y] (ab76149)

    YB1 was immunoprecipitated using 0.5mg HEK293 whole cell extract, 10μg of Rabbit monoclonal to YB1 and 50μl 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 40μl SDS loading buffer and incubated for 10min at 70°C; 10μl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with unpurified ab76149.
    Secondary: Mouse monoclonal [SB62a] secondary antibody to rabbit IgG light chain (HRP) (Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) ab99697).
    Band: 46kDa: YB1.

    All lanes: Immunoprecipitation - Anti-YB1 antibody [EP2708Y] (ab76149)

    Predicted band size: 36 kDa

  • Western blot - Anti-YB1 antibody [EP2708Y] (ab76149), expandable thumbnail

    Western blot - Anti-YB1 antibody [EP2708Y] (ab76149)

    All lanes: Western blot - Anti-YB1 antibody [EP2708Y] (ab76149) at 1/200000 dilution

    Lane 1: HeLa cell lysate at 10 µg

    Lane 2: SW480 cell lysate at 10 µg

    Lane 3: A549 cell lysate at 10 µg

    Lane 4: MCF7 cell lysate at 10 µg

    Secondary

    All lanes: HRP-conjugated goat anti-rabbit IgG at 1/1000 dilution

    Predicted band size: 36 kDa

    Observed band size: 50 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YB1 antibody [EP2708Y] (ab76149), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YB1 antibody [EP2708Y] (ab76149)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling YB1 with unpurified ab76149 at a dilution of 1/100.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Flow Cytometry (Intracellular) - Anti-YB1 antibody [EP2708Y] (ab76149), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-YB1 antibody [EP2708Y] (ab76149)

    Overlay histogram showing HeLa cells stained with unpurified ab76149 (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 (unpurified ab76149, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1μg/1x106 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.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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