Product Info Summary
SKU: | PB9465 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | Flow Cytometry, IF, IHC, IHC-F, ICC, WB |
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Product info
Product Name
Anti-YB1/YBX1 Antibody Picoband™
SKU/Catalog Number
PB9465
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-YB1/YBX1 Antibody Picoband™ catalog # PB9465. Tested in Flow Cytometry, IF, IHC, IHC-F, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-YB1/YBX1 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9465)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human YB1, identical to the related mouse and rat sequences.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9465 is reactive to YBX1 in Human, Mouse, Rat
Applications
PB9465 is guaranteed for Flow Cytometry, IF, IHC, IHC-F, ICC, WB Boster Guarantee
Observed Molecular Weight
50 kDa
Calculated molecular weight
35924 MW
Background of YB1
YBX1 (Y box binding protein 1), commonly referred to as "YB-1" by researchers, is a human protein. YB1 binding has an absolute requirement for the CCAAT box and relative specificity for the Y box. It has a molecular mass of 35,414 and contains 18% basic residues and putative nuclear localization signals. The YBX1 gene is mapped on 1p34.2. Ybx1 was highly expressed in mouse erythroid myeloid lymphoid clone-1 (EML), a hematopoietic precursor cell line, but that it was downregulated in myeloid progenitors and in Gmcsf-treated EML cells during myeloid differentiation. Ybx1 was expressed at high levels in myeloid leukemic cells at different developmental stages. Knockdown of YBX1 in a human leukemic cell line inhibited proliferation ability, induced apoptosis, and induced megakaryocytic differentiation in response to arsenic trioxide treatment. YBX1 is downregulated during myeloid differentiation and aberrant YBX1 expression in leukemic cells may contribute to leukemia development by blocking differentiation.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Assay dilution & Images
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Immunohistochemistry (Frozen Section), 0.5-1μg/ml, Human, -
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry, 1-3μg/1x106 cells, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of YB1 using anti-YB1 antibody (PB9465).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human Jurkat whole cell lysates,
Lane 4: human T47D whole cell lysates,
Lane 5: rat spleen tissue lysates,
Lane 6: mouse RAW264.7 whole cell lysates.
red to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-YB1 antigen affinity purified polyclonal antibody (Catalog # PB9465) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for YB1 at approximately 50 kDa. The expected band size for YB1 is at 36 kDa.
Click image to see more details
Figure 2. IHC analysis of YB using anti-YB antibody (PB9465).
YB was detected in paraffin-embedded section of mouse testis tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-YB Antibody (PB9465) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of YB using anti-YB antibody (PB9465).
YB was detected in paraffin-embedded section of rat testis tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-YB Antibody (PB9465) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of YB using anti-YB antibody (PB9465).
YB was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-YB Antibody (PB9465) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 5. IHC analysis of YB using anti-YB antibody (PB9465).
YB was detected in frozen section of human placenta tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-YB Antibody (PB9465) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 6. IF analysis of YB1 using anti-YB1 antibody (PB9465).
YB1 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-YB1 Antibody (PB9465) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 7. Flow Cytometry analysis of A431 cells using anti-YB1 antibody (PB9465).
Overlay histogram showing A431 cells stained with PB9465 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-YB1 Antibody (PB9465, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For YBX1 (Source: Uniprot.org, NCBI)
Gene Name
YBX1
Full Name
Y-box-binding protein 1
Weight
35924 MW
Superfamily
YBX1 family
Alternative Names
BP-8; CBF-A; class II, Y box-binding protein I; CSDA2; CSDB; DBPB CCAAT-binding transcription factor I subunit A; EFI-A; Enhancer factor I subunit A; MDR-NF1; MGC104858; MGC110976; MGC117250; NSEP1 Y-box-binding protein 1; nuclease-sensitive element-binding protein 1; Y box binding protein 1; YB1 DNA-binding protein B; YB-1 Y-box transcription factor; YBX1 YBX1 BP-8, CBF-A, CSDA2, CSDB, DBPB, EFI-A, MDR-NF1, NSEP-1, NSEP1, YB-1, YB1 Y-box binding protein 1 Y-box-binding protein 1|CCAAT-binding transcription factor I subunit A|DNA-binding protein B|Y-box transcription factor|enhancer factor I subunit A|major histocompatibility complex, class II, Y box-binding protein I|nuclease-sensitive element-binding protein 1
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on YBX1, check out the YBX1 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for YBX1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-YB1/YBX1 Antibody Picoband™ (PB9465)
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Customer Q&As
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6 Customer Q&As for Anti-YB1/YBX1 Antibody Picoband™
Question
Does PB9465 anti-YB1/YBX1 antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2020-03-25
Answer
As indicated on the product datasheet, PB9465 anti-YB1/YBX1 antibody as been tested on IHC. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2020-03-25
Question
I was wanting to use your anti-YB1/YBX1 antibody for IHC for mouse eye on frozen tissues, but I want to know if it has been validated for this particular application. Has this antibody been validated and is this antibody a good choice for mouse eye identification?
Verified Customer
Verified customer
Asked: 2020-03-20
Answer
As indicated on the product datasheet, PB9465 anti-YB1/YBX1 antibody has been tested for IHC, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in mouse eye in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2020-03-20
Question
We are currently using anti-YB1/YBX1 antibody PB9465 for mouse tissue, and we are satisfied with the IHC results. The species of reactivity given in the datasheet says human, mouse, rat. Is it likely that the antibody can work on monkey tissues as well?
J. Williams
Verified customer
Asked: 2019-02-20
Answer
The anti-YB1/YBX1 antibody (PB9465) has not been tested for cross reactivity specifically with monkey tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in monkey you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-02-20
Question
Please see the WB image, lot number and protocol we used for eye using anti-YB1/YBX1 antibody PB9465. Please let me know if you require anything else.
S. Miller
Verified customer
Asked: 2017-06-02
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2017-06-02
Question
We need to test anti-YB1/YBX1 antibody PB9465 on mouse eye for research purposes, then I may be interested in using anti-YB1/YBX1 antibody PB9465 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
O. Jackson
Verified customer
Asked: 2017-05-30
Answer
The products we sell, including anti-YB1/YBX1 antibody PB9465, are only intended for research use. They would not be suitable for use in diagnostic work. If you have the means to develop a product into diagnostic use, and are interested in collaborating with us and develop our product into an IVD product, please contact us for more discussions.
Boster Scientific Support
Answered: 2017-05-30
Question
I see that the anti-YB1/YBX1 antibody PB9465 works with IHC, what is the protocol used to produce the result images on the product page?
B. Williams
Verified customer
Asked: 2015-07-30
Answer
You can find protocols for IHC on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to [email protected]
Boster Scientific Support
Answered: 2015-07-30