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Product Info Summary
| SKU: | M00228-1 |
|---|---|
| Size: | 100µg |
| Reactive Species: | Human |
| Host: | Mouse |
| Application: | IF, IHC, WB |
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Product info
Product Name
Anti-Nucleolin Monoclonal Antibody (4E2)
SKU/Catalog Number
M00228-1
Size
100µg
Form
Liquid. In PBS containing 50% glycerol.
Description
Boster Bio Anti-Nucleolin Monoclonal Antibody (4E2) catalog # M00228-1. Tested in IF, IHC, WB applications. This antibody reacts with Human.
Storage & Handling
Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-Nucleolin Monoclonal Antibody (4E2) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00228-1)
Host
Mouse
Contents
Liquid. In PBS containing 50% glycerol.
Clonality
Monoclonal
Clone Number
4E2
Isotype
IgG1, kappa
Immunogen
Human nucleolin from Raji cell extract.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Reactive Species
M00228-1 is reactive to NCL in Human
Reconstitution
Restore with deionized water (or equivalent) for reconstitution volume of 100 µL
Calculated molecular weight
37682 MW
Background of Nucleolin
Recognizes a protein of 70kDa, which is identified as CD86 (HLDA V; WS Code BP BP072. HLDA V; WS Code A A109. HLDA VI; WS Code BP 95. HLDA VI; WS Code B CD86.9). CD86 is a type I transmembrane glycoprotein and a member of the immunoglobulin superfamily of cell surface receptors. It is expressed at high levels on resting peripheral monocytes and dendritic cells and at very low density on resting B and T lymphocytes. CD86 expression is rapidly upregulated by B cell specific stimuli with peak expression at 18 to 42 hours after stimulation. CD86, along with CD80/B71, is an important accessory molecule in T cell co-stimulation via its interaction with CD28 and CD152/CTLA4. Since CD86 has rapid kinetics of induction, it is believed to be the major CD28 ligand expressed early in the immune response. It is also found on malignant Hodgkin and Reed Sternberg (HRS) cells in Hodgkin's disease.
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
M00228-1 is guaranteed for IF, IHC, 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.
Flow Cytometry (5-10µg/ml final concentration)
Immunocytochemistry (1-10µg/ml)
Immunohistochemistry (1-10µg/ml)
Western Blot (1µg/ml, ECL)
Suggested dilutions/conditions may not be available for all applications.
Optimal conditions must be determined individually for each application.
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of NCL using anti-NCL antibody (M00228-1).
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 50ug of sample under reducing conditions.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-NCL antigen affinity purified polyclonal antibody (Catalog # M00228-1) at 0.5 ug/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-Mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # SA1021) with Tanon 5200 system. A specific band was detected for NCL.
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Figure 2. IHC analysis of NCL using anti-NCL antibody (M00228-1).
NCL was detected in paraffin-embedded section. 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 1ug/ml rabbit anti-NCL Antibody (M00228-1) overnight at 4°C. Biotinylated goat anti Mouse IgG antibody was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Protein Target Info & Infographic
Gene/Protein Information For NCL (Source: Uniprot.org, NCBI)
Gene Name
NCL
Full Name
Nucleolin
Weight
37682 MW
Alternative Names
T-lymphocyte activation antigen CD86;Activation B7-2 antigen;B70;BU63;CTLA-4 counter-receptor B7.2;FUN-1;CD86;CD86;CD28LG2; NCL C23, Nsr1 nucleolin nucleolin
*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 NCL, check out the NCL 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 NCL: 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-Nucleolin Monoclonal Antibody (4E2) (M00228-1)
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Customer Q&As
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4 Customer Q&As for Anti-Nucleolin Monoclonal Antibody (4E2)
Question
We were happy with the WB result of your anti-Nucleolin Monoclonal antibody (4E2) . However we have been able to see positive staining in lymphoma nucleolus. cytoplasm. using this antibody. Is that expected? Could you tell me where is NCL supposed to be expressed?
Verified Customer
Verified customer
Asked: 2019-09-27
Answer
According to literature, lymphoma does express NCL. Generally NCL expresses in nucleus, nucleolus. cytoplasm. Regarding which tissues have NCL expression, here are a few articles citing expression in various tissues:
Cervix carcinoma, Pubmed ID: 8321232, 12429849, 16964243, 17081983, 17924679, 18669648, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Colon carcinoma, Pubmed ID: 24129315
Hair follicle dermal papilla, Pubmed ID: 14702039
Leukemic T-cell, Pubmed ID: 19690332
Liver, Pubmed ID: 24275569
Lymphoblast, Pubmed ID: 14654843
Lymphoma, Pubmed ID: 7718615
Prostate cancer, Pubmed ID: 17487921
Retina, Pubmed ID: 2737305
Boster Scientific Support
Answered: 2019-09-27
Question
We have tried in the past anti-Nucleolin Monoclonal antibody (4E2) for WB on leukemic t-cell in the past. I am using human, and I plan to use the antibody for IHC next. I am interested in examining leukemic t-cell as well as hair follicle dermal papilla in our next experiment. Could you please give me some suggestion on which antibody would work the best for IHC?
P. Williams
Verified customer
Asked: 2019-03-11
Answer
I looked at the website and datasheets of our anti-Nucleolin Monoclonal antibody (4E2) and it appears that M00228-1 has been validated on human in both WB and IHC. Thus M00228-1 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: 2019-03-11
Question
We have seen staining in human colon carcinoma. Are there any suggestions? Is anti-Nucleolin Monoclonal antibody (4E2) supposed to stain colon carcinoma positively?
Verified Customer
Verified customer
Asked: 2018-05-10
Answer
According to literature colon carcinoma does express NCL. According to Uniprot.org, NCL is expressed in testis, retina, hair follicle dermal papilla, lymphoma, cervix carcinoma, lymphoblast, prostate cancer, leukemic t-cell, cervix carcinoma erythroleukemia, liver, colon carcinoma, among other tissues. Regarding which tissues have NCL expression, here are a few articles citing expression in various tissues:
Cervix carcinoma, Pubmed ID: 8321232, 12429849, 16964243, 17081983, 17924679, 18669648, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Colon carcinoma, Pubmed ID: 24129315
Hair follicle dermal papilla, Pubmed ID: 14702039
Leukemic T-cell, Pubmed ID: 19690332
Liver, Pubmed ID: 24275569
Lymphoblast, Pubmed ID: 14654843
Lymphoma, Pubmed ID: 7718615
Prostate cancer, Pubmed ID: 17487921
Retina, Pubmed ID: 2737305
Boster Scientific Support
Answered: 2018-05-10
Question
We are currently using anti-Nucleolin Monoclonal antibody (4E2) M00228-1 for human tissue, and we are well pleased with the IHC results. The species of reactivity given in the datasheet says human. Is it true that the antibody can work on goat tissues as well?
B. Parker
Verified customer
Asked: 2013-02-22
Answer
The anti-Nucleolin Monoclonal antibody (4E2) (M00228-1) has not been validated for cross reactivity specifically with goat tissues, but 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 goat you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2013-02-22
