Fibrillarin Antibody (38F3) 0.25 ml Nucleolar Marker

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Western Blot: Fibrillarin Antibody (38F3) [NB300-269] - Western blot analysis of Fibrillarin expression on yeast protein extracts using NB300-269.
Immunocytochemistry/Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - IF Confocal analysis of MCF7 cells using Fibrillarin antibody (NB300-269, 1:5). An Alexa Fluor 488-conjugated Goat to mouse IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).
Immunohistochemistry-Paraffin: Fibrillarin Antibody (38F3) [NB300-269] - IHC staining of Fibrillarin in human ovarian cancer using DAB with hematoxylin counterstain.
Flow Cytometry: Fibrillarin Antibody (38F3) [NB300-269] - Fibrillarin antibody was tested at 1:400 in HEK293 cells using an Alexa Fluor 488 secondary (shown in green) alongside unstained cells (shown in red).
Immunocytochemistry/Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - Human SH-SY5Y cells stained with NB300-269, showing prominent specular nucleolar staining. The nuclei are counter stained with DAPI (blue), so these spots appear very pale blue.  Neurofilament heavy protein was stained with NB300-217 (red).
Immunocytochemistry/Immunofluorescence: Fibrillarin Antibody (38F3) [NB300-269] - Fibrillarin immunofluorescence in fibroblasts. Image from verified customer review.
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Ordering Information: NB300-269

  • Catalog Number
    NB300-269
  • Displayed Format
    Unconjugated
  • Size(s)
    0.25 ml
  • Availability
  • Price
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Fibrillarin Antibody (38F3) Summary

Species Human, Mouse, Rat, C. elegans, Chicken, Drosophila, Plant, Yeast
Tested Applications WB, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IHC-WhMt
Clonality
Monoclonal
Host
Mouse
Gene
FBL
Purity
Tissue culture supernatant
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Fibrillarin Antibody (38F3) Details

Clone
38F3
Isotype
IgG1
Marker
Nucleolar Marker
Immunogen
Yeast nuclear preparation (S. cerevisiae).
Localization
Nucleus > nucleolus. Note: Fibrillar region of the nucleolus.

Species Reactivity

Human, rat, mouse, chicken, C. elegans, Drosophila, plants and yeast. Other species not tested.
Reviewed Species Read 4 rated 5
using NB300-269 in the following species:

Publications Read Publications using NB300-269 in the following species:


Applications/Dilutions

Dilutions
  • Western Blot 1:1000-1:5000
  • Flow Cytometry 1:20-1:400
  • Immunocytochemistry/Immunofluorescence 1:500-1:1000
  • Immunohistochemistry 1:500-1:1000
  • Immunohistochemistry-Frozen 1:500-1:1000
  • Immunohistochemistry-Paraffin 1:500-1:1000
  • Immunohistochemistry Whole-Mount
Application Notes
This Fibrillarin Antibody (38F3) is useful for Western blot, Immunocytochemistry/Immunofluorescence, Flow Cytometry and Immunohistochemistry on both paraffin-embedded and frozen sections. In WB a band can be seen at approximately 34 kDa. In ICC/IF, this antibody shows prominent specular nucleolar staining. Use in Immunohistochemistry whole mount reported in scientific literature (PMID 24722283)
Molecular Weight
34 kDa
Positive Controls
Reviewed Applications
Read 4 Review rated 5
using
NB300-269 in the following applications:

Publications
Read Publications using
NB300-269 in the following applications:

Contact Information

Product PDFs

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Reviews

Average Rating: 5
Recent Reviews
ICC/IF Tested In Human
Reviewed On: 06/09/2014
Rated: 5

Fibrillarin NB300-269
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Publications

Fibrillarin Antibody (NB300-269) has been mentioned in at least 5 publications.
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Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Unit Size
0.25 ml
Concentration
This product is unpurified. The exact concentration of antibody is not quantifiable.
Preservative
10mM Sodium Azide
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

Gene Symbol FBL
Entrez

Alternate Names for Fibrillarin Antibody (38F3)

  • EC 2.1.1,34-kD nucleolar scleroderma antigen
  • EC 2.1.1.-
  • EC 2.1.1.37
  • FIB
  • FIB1,34 kDa nucleolar scleroderma antigen
  • fibrillarin
  • FLRNrRNA 2'-O-methyltransferase fibrillarin
  • RNA, U3 small nucleolar interacting protein 1
  • RNU3IP1
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Related Products by Gene

Background

Fibrillarin / Nop1p, was originally identified as a nucleolar protein of baker's yeast, Saccharomyces cerevisiae (accession P15646). The Nop1p protein is essential for yeast viability and is localized in the nucleoli. The human homologue of Nop1p is Fibrillarin (accession P22087), a component of the nucleolar small nuclear ribonucleoprotein (snRNP) particle. The human Fibrillarin gene is located on chromosome 19 (19q13.1). Fibrillarin proteins have been cloned and sequenced from several other species (mouse, accession P35550, Xenopus accession P22232, C. elegans accession Q22053, and S. pombe accession P35551). The N terminal 80 amino acids contain multiple copies based on the peptide RGG, and the remaining 240 amino acids consist of the Fibrillarin domain. A Fibrillarin homologue has also been identified in the genome of the archean Methanococcus (accession NC_000909). This protein lacks the RGG rich N-terminal extension but is clearly homologous to the other sequences throughout the entire Fibrillarin domain. The structure of this molecule has been determined and shown to consist of 2 extended beta-sheets flanked by 4 alpha-helixes. Patients with the autoimmune disease scleroderma often have strong circulating autoantibodies to a 34kDa protein which was subsequently found to be Fibrillarin. Fibrillarin is an excellent marker for the nucleolus.

Reviews for Fibrillarin Antibody (NB300-269) (4) 54

Read what people are saying who have used Fibrillarin Antibody (NB300-269).

We have 4 reviews tested in 1 species: Human.
We have 4 reviews tested in 3: Immunocytochemistry/Immunofluorescence, Immunocytochemistry, Immunofluorescence.
Average Rating: 5
(Based on 4 reviews)

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Images Ratings Applications Species Reviewed By Date Details
Immunocytochemistry/Immunofluorescence Fibrillarin NB300-269
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Excellent
ICC/IF Human 06/09/2014
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Summary

ApplicationImmunocytochemistry/Immunofluorescence
SpeciesHuman
Special ApplicationsIF Confocal analysis of MCF7 cells using Fibrillarin antibody (NB300-269, 1:5). An Alexa Fluor 488-conjugated Goat to mouse IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).
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Immunocytochemistry Fibrillarin NB300-269
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Excellent
ICC/IF Human 06/09/2014
Show

Summary

ApplicationImmunocytochemistry
SpeciesHuman
Special ApplicationsIF Confocal analysis of MCF7 cells using Fibrillarin antibody (NB300-269, 1:5). An Alexa Fluor 488-conjugated Goat to mouse IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).
FileView PDF
Immunocytochemistry/Immunofluorescence Fibrillarin NB300-269
Click to Enlarge
Excellent
ICC/IF Human 04/24/2014
Show

Summary

ApplicationImmunocytochemistry/Immunofluorescence
Sample TestedFibroblasts
SpeciesHuman
Lot92412

Blocking

Blocking Detailsblocked in 5% Normal Goat serum/0.3% Triton X-100 in PBS, 1 hour, room temperature

Primary Anitbody

Dilution Ratio1:500, 4C, overnight, in 1X PBS/1% BSA/0.3% Triton X-100

Secondary Antibody

Secondary Manufacturer Cat#cell signaling 4408
Secondary Concentration1:1000

Details

Fixation DetailsFixed in PFA, Blocked in Normal Goat serum containing Triton X-100, Incubated overnight 1:100 in 1% BSA
Wash Description1X PBS, 3 washes 5 minutes each
Immunofluorescence Fibrillarin NB300-269
Click to Enlarge
Excellent
ICC/IF Human 04/24/2014
Show

Summary

ApplicationImmunofluorescence
Sample TestedFibroblasts
SpeciesHuman
Lot92412

Blocking

Blocking Detailsblocked in 5% Normal Goat serum/0.3% Triton X-100 in PBS, 1 hour, room temperature

Primary Anitbody

Dilution Ratio1:500, 4C, overnight, in 1X PBS/1% BSA/0.3% Triton X-100

Secondary Antibody

Secondary Manufacturer Cat#cell signaling 4408
Secondary Concentration1:1000

Details

Fixation DetailsFixed in PFA, Blocked in Normal Goat serum containing Triton X-100, Incubated overnight 1:100 in 1% BSA
Wash Description1X PBS, 3 washes 5 minutes each

Publications for Fibrillarin Antibody (NB300-269) (5)

Read what people are writing who have used Fibrillarin Antibody (NB300-269).

We have publications tested in 4 confirmed species: Human, Mouse, C. elegans, Drosophila.

We have publications tested in 6 applications: WB, ICC/IF, IHC, IHC-Fr, IHC-P, IHC-WhMt.

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Product General Protocols

View specific protocols for Fibrillarin Antibody (NB300-269): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

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FAQs for Fibrillarin (31)

  1. In the immunohistochemistry paraffin section protocol....the PBS buffer....should it be at certain pH?
    • For IHC application, different labs cites the pH of PBS buffer in the range of 7.1 - 7.6 but most commonly used pH is pH 7.4 (this is what we use in our lab). You may use this PBS (pH 7.4) for making permeablization buffer, antibody diluent buffers and for making wash buffer. For more on the protocol that we use in our lab and for IHC-P troubleshooting suggestions, you may visit: IHC-P Protocol and IHC-P Troubleshooting.
  2. I was wondering what does Format 7C mean as far as anitbodies go?
    • 7C is a fluorophore - it is useful for FLOW. The details of the fluor are (A=425, E=500)
  3. What epitope does this monoclonal antibody recognize?
    • R&D Systems does not epitope map its antibodies. The antibody is generated using the entire immunogen listed on the datasheet. If a peptide immunogen is used, this will be indicated on the datasheet.
  4. What epitope does this polyclonal antibody recognize?
    • Polyclonal antibodies generally recognize multiple epitopes because they are generated using the entire immunogen stated on the product datasheet.
  5. What are R&D Systems matched antibody pairs?
    • R&D Systems matched antibody pairs assist in the development of an ELISA. R&D Systems has demonstrated that the paired antibodies can be used in a sandwich immunoassay to recognize the recombinant protein. We recommend that the investigator have expertise in immunoassay development before attempting to use these products. Each investigator must empirically determine the optimal concentrations for the capture and detection antibodies. The amount of antibodies supplied are not normalized to develop the same number of plates. R&D Systems recombinant cytokines are not calibrated by ELISA and therefore must be mass calibrated before use. The DuoSet ELISA Development Kit product line includes the antibodies, standard, enzyme, and protocol necessary for running an immunoassay, and can often be a better value than purchasing antibody pairs and protein standard.
  6. Can I use your antibodies for a matched pair ELISA with a standard from another company?
    • Yes, but there are potential issues to consider. There can be differences in immunological recognition or the binding between antibody and recombinant protein. This can happen if there is a difference in the folding or sequence of the recombinant protein used as the standard versus the recombinant protein used as the immunogen for the antibody. Protein folding or sequence differences in the antibody-binding region can lead to poor (or no) recognition of the standard.
  7. Can I use your antibodies for a matched pair ELISA with a standard from another company?
    • Yes, but there are potential issues to consider. There can be differences in immunological recognition or the binding between antibody and recombinant protein. This can happen if there is a difference in the folding or sequence of the recombinant protein used as the standard versus the recombinant protein used as the immunogen for the antibody. Protein folding or sequence differences in the antibody-binding region can lead to poor (or no) recognition of the standard.
  8. What grade BSA is recommend for use in ELISA and ELISpot Development Systems?
    • The grade of the BSA used has been found to be a critical component for running a successful ELISA. BSA with a minimum purity of 98% is recommended. R&D Systems recommends using the same BSAs used in development of the DuoSet; Catalog # DY995 Reagent Diluent Concentrate 2, Catalog # DY997 Reagent Diluent Concentrate 1. Millipore Probumin® Diagnostic Grade BSA (Millipore, Catalog # 82-045) is another example.
  9. What grade BSA is recommend for use in ELISA and ELISpot Development Systems?
    • The grade of the BSA used has been found to be a critical component for running a successful ELISA. BSA with a minimum purity of 98% is recommended. R&D Systems recommends using the same BSAs used in development of the DuoSet; Catalog # DY995 Reagent Diluent Concentrate 2, Catalog # DY997 Reagent Diluent Concentrate 1. Millipore Probumin® Diagnostic Grade BSA (Millipore, Catalog # 82-045) is another example.
  10. What is a direct ELISA?
    • In a direct ELISA, a plate is coated with the analyte of interest and a labeled detection antibody is used to verify the presence of the analyte.The direct ELISA may use a colorimetric, chemiluminescent, or fluorescent reporter.
  11. What is a direct ELISA?
    • In a direct ELISA, a plate is coated with the analyte of interest and a labeled detection antibody is used to verify the presence of the analyte.The direct ELISA may use a colorimetric, chemiluminescent, or fluorescent reporter.
  12. What is a sandwich ELISA?
    • A sandwich ELISA uses an immobilized capture antibody specific for the analyte of interest in a sample. After the analyte is bound to the immobilized antibody, a labeled secondary antibody specific for the analyte is used for detection. The analyte is "sandwiched" between the two antibodies. The sandwich ELISA is extremely sensitive, and the values obtained are quantitative when compared with a standard curve.
  13. What is a sandwich ELISA?
    • A sandwich ELISA uses an immobilized capture antibody specific for the analyte of interest in a sample. After the analyte is bound to the immobilized antibody, a labeled secondary antibody specific for the analyte is used for detection. The analyte is "sandwiched" between the two antibodies. The sandwich ELISA is extremely sensitive, and the values obtained are quantitative when compared with a standard curve.
  14. What is the difference between AB###, AF###, BAF###, MAB###, and BAM### antibody catalog number designations?
    • AB### designated antibodies are polyclonal antibodies that are purified using Protein A, Protein G or Ion Exchange chromatography. Consequently, the resulting purified antibody is a total IgG fraction and may contain IgG not specific for the analyte of interest. AF### designated antibodies, are antigen affinity-purified and then further purified by ion exchange or size exclusion. Therefore, AF### antibodies are IgG specific only to the antigen. Antibodies that have the designation MAB### are monoclonal antibodies. BAF### and BAM### designated antibodies are the biotinylated versions of the AF### and MAB### designated antibodies, respectively.
  15. What is the difference between AB###, AF###, BAF###, MAB###, and BAM### antibody catalog number designations?
    • AB### designated antibodies are polyclonal antibodies that are purified using Protein A, Protein G or Ion Exchange chromatography. Consequently, the resulting purified antibody is a total IgG fraction and may contain IgG not specific for the analyte of interest. AF### designated antibodies, are antigen affinity-purified and then further purified by ion exchange or size exclusion. Therefore, AF### antibodies are IgG specific only to the antigen. Antibodies that have the designation MAB### are monoclonal antibodies. BAF### and BAM### designated antibodies are the biotinylated versions of the AF### and MAB### designated antibodies, respectively.
  16. What is the rationale for using trehalose to stabilize proteins?
    • Trehalose is an effective sugar for stabilizing proteins against damage caused by freezing. It can also make the protein more resistant to moisture gain when lyophilized, resulting in a product that is less likely to precipitate when reconstituted. In addition, it has been used in approved parenteral therapeutics
  17. What is the rationale for using trehalose to stabilize proteins?
    • Trehalose is an effective sugar for stabilizing proteins against damage caused by freezing. It can also make the protein more resistant to moisture gain when lyophilized, resulting in a product that is less likely to precipitate when reconstituted. In addition, it has been used in approved parenteral therapeutics
  18. Will trehalose affect my conjugation reaction?
    • It is possible that the presence of trehalose will interfere in the successful conjugation of a protein. This will depend on the method used, and the customer should investigate this potential prior to purchasing the product.
  19. Will trehalose affect my conjugation reaction?
    • It is possible that the presence of trehalose will interfere in the successful conjugation of a protein. This will depend on the method used, and the customer should investigate this potential prior to purchasing the product.
  20. Will trehalose affect the performance of the protein or antibody in my specific application?
    • We have seen no adverse effect in our bioassays or other approved applications. However, customers are advised to run a control in their assay to determine if the concentration of trelahose in the protein or antibody formulation has any adverse effects.
  21. Will trehalose affect the performance of the protein or antibody in my specific application?
    • We have seen no adverse effect in our bioassays or other approved applications. However, customers are advised to run a control in their assay to determine if the concentration of trelahose in the protein or antibody formulation has any adverse effects.
  22. Will trehalose included in the formulation affect the animal if it is injected?
    • Trehalose is unlikely to have an effect in vivo. It has been approved as an excipient for use in human injectable drugs.The trehalose used by R&D Systems is derived from Saccharomyces cerevisiae and is determined to be at minimum 98.5% pure by HPAE.
  23. Will trehalose included in the formulation affect the animal if it is injected?
    • Trehalose is unlikely to have an effect in vivo. It has been approved as an excipient for use in human injectable drugs.The trehalose used by R&D Systems is derived from Saccharomyces cerevisiae and is determined to be at minimum 98.5% pure by HPAE.
  24. What are R&D Systems matched antibody pairs?
    • R&D Systems matched antibody pairs assist in the development of an ELISA. R&D Systems has demonstrated that the paired antibodies can be used in a sandwich immunoassay to recognize the recombinant protein. We recommend that the investigator have expertise in immunoassay development before attempting to use these products. Each investigator must empirically determine the optimal concentrations for the capture and detection antibodies. The amount of antibodies supplied are not normalized to develop the same number of plates. R&D Systems recombinant cytokines are not calibrated by ELISA and therefore must be mass calibrated before use. The DuoSet ELISA Development Kit product line includes the antibodies, standard, enzyme, and protocol necessary for running an immunoassay, and can often be a better value than purchasing antibody pairs and protein standard.
  25. What is the recommended protocol for reconstituting and aliquotting lyophilized proteins and antibodies?<br>
    • Following these guidelines for reconstituting lyophilized material will help ensure complete recovery of the protein or antibody.

      1) Tap or briefly centrifuge the vial before opening to dislodge any lyophilized material that may be dispersed on the wall or cap of the vial.

      2) Use the buffer and stock concentration recommended in the product datasheet. If you want a stock concentration that is higher than the one recommended, contact Technical Service for specific recommendations.

      3) For optimal recovery, reconstitute using room temperature buffer.

      4) After adding the buffer, re-cap the vial and invert gently by hand or place on a slow rocking platform. This will allow the reconstitution buffer to coat all the surfaces inside the vial. Do not mix by vortexing or by pipetting the material up and down.

      5) Allow the vial to sit at room temperature with gentle agitation for at least 15 minutes before aliquotting or using.

      6) Store the reconstituted protein in polypropylene or siliconized tubes. If aliquotting, it is recommended that aliquots be no smaller than 10 μL. In addition, avoid repeated freeze/thaw cycles.

  26. What is the recommended protocol for reconstituting and aliquotting lyophilized proteins and antibodies?<br>
    • Following these guidelines for reconstituting lyophilized material will help ensure complete recovery of the protein or antibody.

      1) Tap or briefly centrifuge the vial before opening to dislodge any lyophilized material that may be dispersed on the wall or cap of the vial.

      2) Use the buffer and stock concentration recommended in the product datasheet. If you want a stock concentration that is higher than the one recommended, contact Technical Service for specific recommendations.

      3) For optimal recovery, reconstitute using room temperature buffer.

      4) After adding the buffer, re-cap the vial and invert gently by hand or place on a slow rocking platform. This will allow the reconstitution buffer to coat all the surfaces inside the vial. Do not mix by vortexing or by pipetting the material up and down.

      5) Allow the vial to sit at room temperature with gentle agitation for at least 15 minutes before aliquotting or using.

      6) Store the reconstituted protein in polypropylene or siliconized tubes. If aliquotting, it is recommended that aliquots be no smaller than 10 μL. In addition, avoid repeated freeze/thaw cycles.

  27. Can this antibody be used on frozen or paraffin-embedded&nbsp; sections?
    • R&D Systems will provide support for an antibody that is validated for the IHC/ICC application in both frozen and paraffin-embedded sections unless otherwise specified on the datasheet. Should the product fail to work after contacting us for assistance, R&D Systems will offer a product credit toward a future purchase.

      If the datasheet does not have an IHC/ICC claim, we do not have any in-house or collaborative data available to support this application and we cannot guarantee results. It is up to the user to decide if they wish to use the antibody in their application.

      Customers may use the Citations tab on the product-specific web page or contact Technical Service to check for references using a product in an application or with a cell type R&D Systems has not tested in-house.
  28. Can this antibody be used on frozen or paraffin-embedded&nbsp; sections?
    • R&D Systems will provide support for an antibody that is validated for the IHC/ICC application in both frozen and paraffin-embedded sections unless otherwise specified on the datasheet. Should the product fail to work after contacting us for assistance, R&D Systems will offer a product credit toward a future purchase.

      If the datasheet does not have an IHC/ICC claim, we do not have any in-house or collaborative data available to support this application and we cannot guarantee results. It is up to the user to decide if they wish to use the antibody in their application.

      Customers may use the Citations tab on the product-specific web page or contact Technical Service to check for references using a product in an application or with a cell type R&D Systems has not tested in-house.
  29. Can this antibody be used on frozen or paraffin-embedded&nbsp; sections?
    • R&D Systems will provide support for an antibody that is validated for the IHC/ICC application in both frozen and paraffin-embedded sections unless otherwise specified on the datasheet. Should the product fail to work after contacting us for assistance, R&D Systems will offer a product credit toward a future purchase.

      If the datasheet does not have an IHC/ICC claim, we do not have any in-house or collaborative data available to support this application and we cannot guarantee results. It is up to the user to decide if they wish to use the antibody in their application.

      Customers may use the Citations tab on the product-specific web page or contact Technical Service to check for references using a product in an application or with a cell type R&D Systems has not tested in-house.
  30. Has a product ever been used for an application that is not listed on the datasheet?
    • If a specific application is not listed on the datasheet, it may mean that this product has not been tested in this application, or it may mean that in-house testing in this application did not meet R&D Systems’specifications. Please check our Citations tab to see if other researchers have published using your application, sample type and/or species. If you would like more information on whether or not an application has been tested, please contact Technical Service at (800) 343-7475.

  31. Has a product ever been used for an application that is not listed on the datasheet?
    • If a specific application is not listed on the datasheet, it may mean that this product has not been tested in this application, or it may mean that in-house testing in this application did not meet R&D Systems’specifications. Please check our Citations tab to see if other researchers have published using your application, sample type and/or species. If you would like more information on whether or not an application has been tested, please contact Technical Service at (800) 343-7475.