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Product Info Summary
| SKU: | PB9798 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-ROC1/RBX1 Antibody Picoband®
SKU/Catalog Number
PB9798
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-ROC1/RBX1 Antibody Picoband® catalog # PB9798. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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-ROC1/RBX1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9798)
Host
Rabbit
Contents
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human ROC1, identical to the related mouse sequence.
*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
PB9798 is reactive to RBX1 in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
15 kDa
Calculated molecular weight
12274 MW
Background of RBX1
RING-box protein 1, also known as ROC1, is a protein that in humans is encoded by the RBX1 gene. This gene is mapped to chromosome 22q13.2 based on an alignment of the RBX1 sequence with the genomic sequence. ROC1 is recruited by cullin-1 to form a quaternary SCF (HOS)-ROC1 holoenzyme (with SKP1 and the BTRCP homolog HOS). SCF (HOS)-ROC1 binds IKK-beta-phosphorylated I-kappa-B-alpha and catalyzes its ubiquitination in the presence of ubiquitin, E1, and CDC34. Conclusively, ROC1 plays a unique role in the ubiquitination reaction by heterodimerizing with cullin-1 to catalyze ubiquitin polymerization.
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.
Application & Images
Applications
PB9798 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee
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, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: Rat Testis Tissue, Rat Brain Tissue, Mouse Brain Tissue, Mouse Spleen Tissue
ICC/IF: Hela cell
FCM: A431 cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of ROC1 using anti-ROC1 antibody (PB9798).
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 50 ug of sample under reducing conditions.
Lane 1: Rat Testis Tissue Lysate,
Lane 2: Rat Brain Tissue Lysate,
Lane 3: Mouse Brain Tissue Lysate,
Lane 4: Mouse Spleen Tissue Lysate.
After electrophoresis, proteins were transferred 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-ROC1 antigen affinity purified polyclonal antibody (Catalog # PB9798) 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 ROC1 at approximately 15 kDa. The expected band size for ROC1 is at 15 kDa.
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Figure 2. Flow Cytometry analysis of A431 cells using anti-ROC1 antibody (PB9798).
Overlay histogram showing A431 cells stained with PB9798 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ROC1 Antibody (PB9798,1μg/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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
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Figure 3. IF analysis of ROC1 using anti-ROC1 antibody (PB9798).
ROC1 was detected in immunocytochemical section of Hela 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 2μg/mL rabbit anti-ROC1 Antibody (PB9798) 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.
Protein Target Info & Infographic
Gene/Protein Information For RBX1 (Source: Uniprot.org, NCBI)
Gene Name
RBX1
Full Name
E3 ubiquitin-protein ligase RBX1
Weight
12274 MW
Superfamily
RING-box family
Alternative Names
E3 ubiquitin-protein ligase RBX1;6.3.2.-;Protein ZYP;RING finger protein 75;RING-box protein 1;Rbx1;Regulator of cullins 1;E3 ubiquitin-protein ligase RBX1, N-terminally processed;RBX1;RNF75, ROC1; RBX1 BA554C12.1, RNF75, ROC1 ring-box 1 E3 ubiquitin-protein ligase RBX1|E3 ubiquitin-protein transferase RBX1|RING finger protein 75|RING-box protein 1|ZYP protein|regulator of cullins 1|ring-box 1, E3 ubiquitin protein ligase
*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 RBX1, check out the RBX1 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 RBX1: 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-ROC1/RBX1 Antibody Picoband® (PB9798)
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Customer Q&As
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5 Customer Q&As for Anti-ROC1/RBX1 Antibody Picoband®
Question
My question regarding product PB9798, anti-ROC1/RBX1 antibody. I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
Verified Customer
Verified customer
Asked: 2019-07-26
Answer
We do not advise storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free PB9798 anti-ROC1/RBX1 antibody, we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2019-07-26
Question
I was wanting to use your anti-ROC1/RBX1 antibody for WB for rat cervix carcinoma 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 rat cervix carcinoma identification?
Verified Customer
Verified customer
Asked: 2019-04-08
Answer
As indicated on the product datasheet, PB9798 anti-ROC1/RBX1 antibody has been validated for WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in rat cervix carcinoma in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2019-04-08
Question
Do you have a BSA free version of anti-ROC1/RBX1 antibody PB9798 available?
Verified Customer
Verified customer
Asked: 2019-02-28
Answer
Thank you for your recent telephone inquiry. I can confirm that some lots of this anti-ROC1/RBX1 antibody PB9798 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2019-02-28
Question
Is a blocking peptide available for product anti-ROC1/RBX1 antibody (PB9798)?
Verified Customer
Verified customer
Asked: 2019-01-17
Answer
We do provide the blocking peptide for product anti-ROC1/RBX1 antibody (PB9798). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2019-01-17
Question
We are currently using anti-ROC1/RBX1 antibody PB9798 for mouse tissue, and we are satisfied with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it likely that the antibody can work on bovine tissues as well?
C. Anderson
Verified customer
Asked: 2017-07-21
Answer
The anti-ROC1/RBX1 antibody (PB9798) has not been tested for cross reactivity specifically with bovine 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 bovine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2017-07-21
