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Product Info Summary
| SKU: | RP1076 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-CD59 glycoprotein CD59 Antibody Picoband®
SKU/Catalog Number
RP1076
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-CD59 glycoprotein CD59 Antibody catalog # RP1076. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human. 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-CD59 glycoprotein CD59 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # RP1076)
Host
Rabbit
Contents
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human CD59 recombinant protein (Position: L26-N102). Human CD59 shares 47.1% amino acid (aa) sequence identity with rat CD59.
*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
RP1076 is reactive to CD59 in Human
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
19 kDa
Calculated molecular weight
14177 MW
Background of CD59
This gene encodes a cell surface glycoprotein that regulates complement-mediated cell lysis, and it is involved in lymphocyte signal transduction. And this protein is a potent inhibitor of the complement membrane attack complex, whereby it binds complement C8 and/or C9 during the assembly of this complex, thereby inhibiting the incorporation of multiple copies of C9 into the complex, which is necessary for osmolytic pore formation. It also plays a role in signal transduction pathways in the activation of T cells. Mutations in this gene cause CD59 deficiency, a disease resulting in hemolytic anemia and thrombosis, and which causes cerebral infarction. Multiple alternatively spliced transcript variants, which encode the same protein, have been identified for this gene.
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
RP1076 is guaranteed for Flow Cytometry, IF, IHC, 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, 0.1-0.25 μg/ml
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, By Heat
Immunocytochemistry/Immunofluorescence, 5μg/ml
Flow Cytometry (Fixed), 1-3μg/1x106cells
Positive Control
WB: human Hela whole cell, human PC-3 whole cell
IHC: human tonsil tissue
ICC/IF: A431 cell
FCM: K562 cell
Validation Images & Assay Conditions
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Figure 1. IHC analysis of CD59 using anti-CD59 antibody (RP1076).
CD59 was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-CD59 Antibody (RP1076) 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.
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Figure 2. Flow Cytometry analysis of K562 cells using anti-CD59 antibody (RP1076).
Overlay histogram showing K562 cells stained with RP1076 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-CD59 Antibody (RP1076,1μg/1x106 cells) for 30 min at 20°C. DyLight488 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. Western blot analysis of CD59 using anti-CD59 antibody (RP1076).
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 PC-3 whole cell lysates.
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-CD59 antigen affinity purified polyclonal antibody (Catalog # RP1076) at 0.25 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 CD59 at approximately 19 kDa. The expected band size for CD59 is at 14 kDa.
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Figure 4. IF analysis of CD59 using anti-CD59 antibody (RP1076).
CD59 was detected in an 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 5 μg/mL rabbit anti-CD59 Antibody (RP1076) 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 CD59 (Source: Uniprot.org, NCBI)
Gene Name
CD59
Full Name
CD59 glycoprotein
Weight
14177 MW
Alternative Names
CD59 glycoprotein;1F5 antigen;20 kDa homologous restriction factor;HRF-20;HRF20;MAC-inhibitory protein;MAC-IP;MEM43 antigen;Membrane attack complex inhibition factor;MACIF;Membrane inhibitor of reactive lysis;MIRL;Protectin;CD59;CD59;MIC11, MIN1, MIN2, MIN3, MSK21; CD59 16.3A5, 1F5, EJ16, EJ30, EL32, G344, HRF-20, HRF20, MAC-IP, MACIF, MEM43, MIC11, MIN1, MIN2, MIN3, MIRL, MSK21, p18-20 CD59 molecule (CD59 blood group) CD59 glycoprotein|1F5 |20 kDa homologous restriction factor|CD59 p18-20 ( by antibodies 16.3A5, EJ16, EJ30, EL32 and G344)|CD59 blood group |CD59 molecule, complement regulatory protein|Ly-6-like protein|MEM43 |T cell-activating protein|human leukocyte MIC11|lymphocytic CD59/MEM43|membrane attack complex (MAC) inhibition factor|membrane attack complex inhibition factor|membrane inhibitor of reactive lysis|protectin|surface anitgen recognized by 16.3A5
*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 CD59, check out the CD59 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 CD59: 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-CD59 glycoprotein CD59 Antibody Picoband® (RP1076)
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Customer Q&As
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5 Customer Q&As for Anti-CD59 glycoprotein CD59 Antibody Picoband®
Question
We need using your anti-CD59 antibody for copii-mediated vesicle transport studies. Has this antibody been tested with western blotting on human tonsil tissue? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2020-02-17
Answer
We appreciate your inquiry. This RP1076 anti-CD59 antibody is tested on human tonsil tissue, k562 cells. It is guaranteed to work for Flow Cytometry, IHC, ICC in human. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2020-02-17
Question
Our lab used your anti-CD59 antibody for ICC on colon in the past. I am using human, and I plan to use the antibody for IHC next. I am looking for examining colon as well as chorionic villus in our next experiment. Could you please give me some suggestion on which antibody would work the best for IHC?
Verified Customer
Verified customer
Asked: 2020-01-24
Answer
I took a look at the website and datasheets of our anti-CD59 antibody and it appears that RP1076 has been tested on human in both ICC and IHC. Thus RP1076 should work for your application. Our Boster satisfaction guarantee will cover this product for IHC in human even if the specific tissue type has not been validated. We do have a comprehensive range of products for IHC detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2020-01-24
Question
We have seen staining in human colon. Any tips? Is anti-CD59 antibody supposed to stain colon positively?
Verified Customer
Verified customer
Asked: 2019-02-28
Answer
According to literature colon does express CD59. According to Uniprot.org, CD59 is expressed in chorionic villus, t-cell, blood, colon, milk, liver, urine, among other tissues. Regarding which tissues have CD59 expression, here are a few articles citing expression in various tissues:
Blood, Pubmed ID: 1383553
Colon, Pubmed ID: 15489334
Liver, Pubmed ID: 24275569
Milk, Pubmed ID: 18780401
T-cell, Pubmed ID: 2475570
Urine, Pubmed ID: 7520819
Boster Scientific Support
Answered: 2019-02-28
Question
My team were content with the WB result of your anti-CD59 antibody. However we have been able to see positive staining in liver cell membrane using this antibody. Is that expected? Could you tell me where is CD59 supposed to be expressed?
C. Edwards
Verified customer
Asked: 2013-11-26
Answer
From literature, liver does express CD59. Generally CD59 expresses in cell membrane. Regarding which tissues have CD59 expression, here are a few articles citing expression in various tissues:
Blood, Pubmed ID: 1383553
Colon, Pubmed ID: 15489334
Liver, Pubmed ID: 24275569
Milk, Pubmed ID: 18780401
T-cell, Pubmed ID: 2475570
Urine, Pubmed ID: 7520819
Boster Scientific Support
Answered: 2013-11-26
Question
We are currently using anti-CD59 antibody RP1076 for human tissue, and we are happy with the ICC results. The species of reactivity given in the datasheet says human. Is it possible that the antibody can work on dog tissues as well?
P. Gonzalez
Verified customer
Asked: 2013-08-12
Answer
The anti-CD59 antibody (RP1076) has not been validated for cross reactivity specifically with dog 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 dog you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2013-08-12
