CD45 Antibody (30-F11) - BSA Free

Novus Biologicals | Catalog # NB100-77417

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Functional, In vitro assay, In vivo assay, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, In vitro assay, In vivo assay, Functional Assay, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG2b Kappa Clone # 30-F11

Format

BSA Free
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Product Specifications

Immunogen

This CD45 Antibody (30-F11) was prepared from Mouse thymus or spleen.

Reactivity Notes

Use in Human reported in scientific literature (PMID:34494385).

Localization

Transmembrane Protein with Extracellular, Helical and Cytoplasmic Domains

Specificity

Recognizes all CD45 isoforms as well as the known mouse CD45 alloantigens (CD45-1/Ly 5a and CD45-2/Ly 5b)

Clonality

Monoclonal

Host

Rat

Isotype

IgG2b Kappa

Theoretical MW

145 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for CD45 Antibody (30-F11) - BSA Free

Flow Cytometry: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

Flow Cytometry: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

Flow Cytometry: CD45 Antibody (30-F11) [NB100-77417] - Analysis of CD45 (30F11) in freshly isolated mouse (Balb C) splenocytes (1x10^6) were stained with [NB100-77417] at 1:1000 dilution and detected with FITC conjugated donkey anti-rat IgG secondary (orange). Shown with rat IgG isotype control (blue).
Immunohistochemistry-Paraffin: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

Immunohistochemistry-Paraffin: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

Immunohistochemistry-Paraffin: CD45 Antibody (30-F11) [NB100-77417] - Analysis of FFPE section of mouse spleen using rat monoclonal CD45 antibody (30-F11) [NB100-77417] at 1:50 dilution. The antibody generated an expected membrane staining in the red and white pulp spleenocytes of the tested spleen section.
Immunohistochemistry-Paraffin: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

Immunohistochemistry-Paraffin: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

Immunohistochemistry-Paraffin: CD45 Antibody (30-F11) [NB100-77417] - Mouse glomeruli stained with CD45 antibody. 4% formaldehyde fixed paraffin sections. EDTA was used for antigen retrieval. IHC-P image submitted by a verified customer review.
Immunohistochemistry: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

Immunohistochemistry: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

Immunohistochemistry: CD45 Antibody (30-F11) [NB100-77417] - Analysis of a FFPE embedded tissue section of mouse liver using 1:100 dilution of CD45 antibody (30-F11) [NB100-77417]. The signal was developed using AEC reagent based method with hematoxylin counterstaining.
Immunohistochemistry-Frozen: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

Immunohistochemistry-Frozen: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

Immunohistochemistry-Frozen: CD45 Antibody (30-F11) [NB100-77417] - Analysis of a mouse spleen cryosection using CD45 antibody (30-F11) [NB100-77417] at 1:50 dilution. The signal was detected using goat anti-rat Alexa fluor 488 secondary with DAPI counterstaining. IHC-P image submitted by a verified customer review.
Immunohistochemistry-Paraffin: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

Immunohistochemistry-Paraffin: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

Immunohistochemistry-Paraffin: CD45 Antibody (30-F11) [NB100-77417] - Staining of a FFPE mouse liver tissue section using 1:100 dilution of CD45 antibody (30-F11) [NB100-77417]. Signal was detected using AF488 conjugated donkey anti-rat secondary antibody at 1:500 dilution. IHC-P image submitted by a verified customer review.
Immunohistochemistry: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

Immunohistochemistry: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

CD45-Antibody-30-F11-Immunohistochemistry-NB100-77417-img0009.jpg
Immunohistochemistry: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

Immunohistochemistry: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

CD45-Antibody-30-F11-Immunohistochemistry-NB100-77417-img0012.jpg
Immunohistochemistry: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

Immunohistochemistry: CD45 Antibody (30-F11) - BSA Free [NB100-77417]

CD45-Antibody-30-F11-Immunohistochemistry-NB100-77417-img0011.jpg

Applications for CD45 Antibody (30-F11) - BSA Free

Application
Recommended Usage

Flow Cytometry

1:10-1:1000

Functional

reported in scientific literature (PMID 7743522)

Immunocytochemistry/ Immunofluorescence

1:10-1:500

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Frozen

1:10-1:500

Immunohistochemistry-Paraffin

1:10-1:500

Immunoprecipitation

1:10-1:500

In vitro assay

reported in scientific literature (PMID 1974916)

In vivo assay

reported in multiple pieces of scientific literature

Western Blot

1:100-1:2000
Application Notes
Each lot of this CD45 antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. IF data from customer review. For immunofluorescent staining, the suggested use of this reagent is: 0.25 microgram per one million cells in 100 microliter volume. It is recommended that the reagent be titrated for optimal performance for each application. This antibody is CyTOF ready. NB100-77417 may also be used for IHC with acetone-fixed frozen sections, zinc-fixed paraffin-embedded sections.

Reviewed Applications

Read 5 reviews rated 4.8 using NB100-77417 in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: CD45

CD45, also known as leukocyte common antigen (LCA), T200, or Ly5, is a member C of the class 1 receptor-like protein tyrosine phosphatase (PTPRC) family (1, 2). It is a transmembrane glycoprotein which, due to alternative splicing, has a multiple isoforms with a theoretical molecular weight ranging from 180 - 220 kDa (1, 3-5). Human CD45 is synthesized as a 1281 amino acid sequence consisting of an alternatively spliced extracellular receptor-like region, a cysteine-rich domain, fibronectin-like III repeats, a transmembrane segment, and a cytoplasmic region with tandem protein tyrosine phosphatase (PTP) domains: the membrane proximal domain (D1) and the membrane distal domain (D2) (3, 5). CD45 is expressed on all nucleated hematopoietic cells and their precursors, except mature red blood cells, and is one of the most abundantly-expressed cell-surface glycoproteins, comprising approximately 10% of surface proteins in lymphocytes (3). Functionally, CD45 is essential for development and activation of T-cells and B-cells (1-5). More specifically, CD45 positively regulates antigen receptor signaling and Src-family member kinase activity (1, 3). There are many ways to regulate CD45 phosphatase activity including ligand binding, dimerization, protein interactions, cellular localization, and covalent modifications (3, 6). Ligands for CD45 include pUL11, a transmembrane protein of the cytomegalovirus RL11 (CMV RL11) family, and placental protein 14 (PP14), both of which exclusively bind CD45, and various lectins including CD22, galectin-1, galectin-3, macrophage mannose receptor (MR), and macrophage galactose-type lectin (MGL) (6).

Given its role in immune cell development and activation, CD45 has also been linked to a variety of diseases. The importance of CD45 in immunity has been revealed in human and mouse studies where CD45-deficiency leads to a severe-combined immunodeficiency (SCID) phenotype (2, 3, 6). A CD45-knockout mice study revealed inhibited thymocyte production and poor B-cell response, whereas CD45 activation in mice causes lymphoproliferation and autoantibody production (3). CD45 variants have been associated with altered immune function and autoimmune disorders including multiple sclerosis, systemic lupus erythematosus (SLE), and rheumatoid arthritis (6). Furthermore, altered CD45 expression has been implicated in oncological conditions including chronic lymphatic leukemia, acute lymphatic leukemia, Hodgkin lymphoma, multiple myeloma, and diffuse large B-cell lymphoma (6). Considering its role in autoimmune disorders, immunodeficiency and cancer, CD45 is an ideal therapeutic target (3, 6). The main approaches to control CD45 function is through either selective inhibitors or anti-CD45 antibodies (3).

Alternative names for CD45 includes B220, CD antigen: CD45, CD45 antigen, CD45R, EC 3.1.3.48, GP180, LCA, Leukocyte common antigen, LY5, protein tyrosine phosphatase receptor type c polypeptide, PTPRC, receptor-type tyrosine-protein phosphatase C, T200 Glycoprotein, and T200.

References

1. Trowbridge, I. S., & Thomas, M. L. (1994). CD45: an emerging role as a protein tyrosine phosphatase required for lymphocyte activation and development. Annual review of immunology. https://doi.org/10.1146/annurev.iy.12.040194.000505

2. Andersen, J. N., Jansen, P. G., Echwald, S. M., Mortensen, O. H., Fukada, T., Del Vecchio, R., Tonks, N. K., & Moller, N. P. (2004). A genomic perspective on protein tyrosine phosphatases: gene structure, pseudogenes, and genetic disease linkage. FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

3. Hermiston, M. L., Xu, Z., & Weiss, A. (2003). CD45: a critical regulator of signaling thresholds in immune cells. Annual review of immunology. https://doi.org/10.1146/annurev.immunol.21.120601.140946

4. Tonks, N. K., Diltz, C. D., & Fischer, E. H. (1990). CD45, an integral membrane protein tyrosine phosphatase. Characterization of enzyme activity. The Journal of biological chemistry.

5. Nam, H. J., Poy, F., Saito, H., & Frederick, C. A. (2005). Structural basis for the function and regulation of the receptor protein tyrosine phosphatase CD45. The Journal of experimental medicine. https://doi.org/10.1084/jem.20041890

6. Rheinlander, A., Schraven, B., & Bommhardt, U. (2018). CD45 in human physiology and clinical medicine. Immunology letters. https://doi.org/10.1016/j.imlet.2018.01.009

Long Name

Cluster of Differentiation 45

Alternate Names

CD45, LCA, PTPRC, T200 Glycoprotein

Entrez Gene IDs

19264 (Mouse)

Gene Symbol

PTPRC

UniProt

Additional CD45 Products

Product Documents for CD45 Antibody (30-F11) - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for CD45 Antibody (30-F11) - BSA Free

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.

Citations for CD45 Antibody (30-F11) - BSA Free

Customer Reviews for CD45 Antibody (30-F11) - BSA Free (5)

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Showing  1 - 5 of 5 reviews Showing All
Filter By:
  • CD45 Antibody (30-F11)
    Name: Anonymous
    Application: Immunohistochemistry-Paraffin
    Sample Tested: glomeruli
    Species: Mouse
    Verified Customer | Posted 10/13/2021
    CD45 Antibody in glomeruli
    4% formaldehyde fixed paraffin sections. EDTA was used for antigen retrieval. 1ry antibody dilution 1:100, 2ry antibody 1:400
    CD45 Antibody (30-F11) - BSA Free NB100-77417
  • CD45 Antibody (30-F11)
    Name: Anonymous
    Application: Immunohistochemistry-Paraffin
    Sample Tested: Mouse spleen cells
    Species: Mouse
    Verified Customer | Posted 09/15/2017
    No permeabilization Antigen retrieval: Citrate buffer, 20 minutes, 90C 1st ab dilution: 1:100
    CD45 Antibody (30-F11) - BSA Free NB100-77417
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: Mouse Spleen
    Species: Rat
    Verified Customer | Posted 11/24/2015
  • Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: mouse liver
    Species: Mouse
    Verified Customer | Posted 10/02/2014
  • Name: Aurora De Ponti
    Application: Immunofluorescence
    Sample Tested: mouse liver
    Species: Mouse
    Verified Customer | Posted 09/05/2014
    CD45 in mouse liver
    CD45 Antibody (30-F11) - BSA Free NB100-77417

There are no reviews that match your criteria.

Showing  1 - 5 of 5 reviews Showing All

Protocols

View specific protocols for CD45 Antibody (30-F11) - BSA Free (NB100-77417):

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for CD45 Antibody (30-F11) - BSA Free

Showing  1 - 4 of 4 FAQs Showing All
  • Q: For use in Western Blot with CD45 antibodies, what molecular weight of the band should I expect to see?

    A: The theoretical molecular weight for most of our CD45 antibodies is 147 kDa based off the first isoform. Any variation on 147 is due to the immunogen being from a different species and the protein being a slightly different size. CD45 is a family of single chain transmembraneous glycoproteins consisting of at least four isoforms (220, 205, 190, 180 kDa) which share a common large intracellular domain. Their extracellular domains are heavily glycosylated.

  • Q: If this product is used in an application or species as a part of a customer review, will that validate this product in the application/species?

    A: If any of our primary antibodes are used in an untested application or species and it is shown to work through images from customer reviews or through publications, this validates the application/species for this product, allowing the tested application/species to fall under our 100% guarantee. Please check out our Innovator's Reward Program if you decide to test a primary antibody with a species or application that is not currently listed. Please note that the Innovator's Reward Program only applies to our primary antibodies.

  • Q: Is CD45 a good target for use in Neurodegeneration studies?

    A: Yes, here are the approved research areas we have listed for our CD45 products: Adaptive Immunity, Cell Biology, Cellular Markers, Cytokine Research, Glia Markers, Hematopoietic Stem Cell Markers, Immunology, Innate Immunity, Mast Cell Markers, Mesenchymal Stem Cell Markers, Microglia Markers, Myeloid Cell Markers, Myeloid-derived Suppressor, Neurodegeneration, Neuroscience, Signal Transduction, Stem Cell Markers, Growth and Development.

  • Q: We would like to order a CD45 antibody to stain immune cells that were isolated from a ligated sciatic nerve of mice with double immunofluorescence (using PFA-fixed tissue on slides). Which of the following rat monoclonal CD45 antibodies would you recommend: 30-F11, IBL-3/16 or 5C16?

    A:

    I would recommend clone 30-F11. As clone IBL-3/16 has not yet been validated for IHC-P application, we would not be able to guarantee its workability on PFA-fixed tissue. Although both other clones i.e. 30-F11 and 5C16, would be good for your samples, CD45 30-F11 # NB100-77417 is a well known clone that offers more flexibility around protocol because of availability of its conjugated forms. It would be advantageous to use a conjugated primary as you are planning for double-immunostaining procedure.

  • Q: For use in Western Blot with CD45 antibodies, what molecular weight of the band should I expect to see?

    A: The theoretical molecular weight for most of our CD45 antibodies is 147 kDa based off the first isoform. Any variation on 147 is due to the immunogen being from a different species and the protein being a slightly different size. CD45 is a family of single chain transmembraneous glycoproteins consisting of at least four isoforms (220, 205, 190, 180 kDa) which share a common large intracellular domain. Their extracellular domains are heavily glycosylated.

  • Q: If this product is used in an application or species as a part of a customer review, will that validate this product in the application/species?

    A: If any of our primary antibodes are used in an untested application or species and it is shown to work through images from customer reviews or through publications, this validates the application/species for this product, allowing the tested application/species to fall under our 100% guarantee. Please check out our Innovator's Reward Program if you decide to test a primary antibody with a species or application that is not currently listed. Please note that the Innovator's Reward Program only applies to our primary antibodies.

  • Q: Is CD45 a good target for use in Neurodegeneration studies?

    A: Yes, here are the approved research areas we have listed for our CD45 products: Adaptive Immunity, Cell Biology, Cellular Markers, Cytokine Research, Glia Markers, Hematopoietic Stem Cell Markers, Immunology, Innate Immunity, Mast Cell Markers, Mesenchymal Stem Cell Markers, Microglia Markers, Myeloid Cell Markers, Myeloid-derived Suppressor, Neurodegeneration, Neuroscience, Signal Transduction, Stem Cell Markers, Growth and Development.

  • Q: We would like to order a CD45 antibody to stain immune cells that were isolated from a ligated sciatic nerve of mice with double immunofluorescence (using PFA-fixed tissue on slides). Which of the following rat monoclonal CD45 antibodies would you recommend: 30-F11, IBL-3/16 or 5C16?

    A:

    I would recommend clone 30-F11. As clone IBL-3/16 has not yet been validated for IHC-P application, we would not be able to guarantee its workability on PFA-fixed tissue. Although both other clones i.e. 30-F11 and 5C16, would be good for your samples, CD45 30-F11 # NB100-77417 is a well known clone that offers more flexibility around protocol because of availability of its conjugated forms. It would be advantageous to use a conjugated primary as you are planning for double-immunostaining procedure.

  • Q: For use in Western Blot with CD45 antibodies, what molecular weight of the band should I expect to see?

    A: The theoretical molecular weight for most of our CD45 antibodies is 147 kDa based off the first isoform. Any variation on 147 is due to the immunogen being from a different species and the protein being a slightly different size. CD45 is a family of single chain transmembraneous glycoproteins consisting of at least four isoforms (220, 205, 190, 180 kDa) which share a common large intracellular domain. Their extracellular domains are heavily glycosylated.

  • Q: If this product is used in an application or species as a part of a customer review, will that validate this product in the application/species?

    A: If any of our primary antibodes are used in an untested application or species and it is shown to work through images from customer reviews or through publications, this validates the application/species for this product, allowing the tested application/species to fall under our 100% guarantee. Please check out our Innovator's Reward Program if you decide to test a primary antibody with a species or application that is not currently listed. Please note that the Innovator's Reward Program only applies to our primary antibodies.

  • Q: Is CD45 a good target for use in Neurodegeneration studies?

    A: Yes, here are the approved research areas we have listed for our CD45 products: Adaptive Immunity, Cell Biology, Cellular Markers, Cytokine Research, Glia Markers, Hematopoietic Stem Cell Markers, Immunology, Innate Immunity, Mast Cell Markers, Mesenchymal Stem Cell Markers, Microglia Markers, Myeloid Cell Markers, Myeloid-derived Suppressor, Neurodegeneration, Neuroscience, Signal Transduction, Stem Cell Markers, Growth and Development.

  • Q: We would like to order a CD45 antibody to stain immune cells that were isolated from a ligated sciatic nerve of mice with double immunofluorescence (using PFA-fixed tissue on slides). Which of the following rat monoclonal CD45 antibodies would you recommend: 30-F11, IBL-3/16 or 5C16?

    A:

    I would recommend clone 30-F11. As clone IBL-3/16 has not yet been validated for IHC-P application, we would not be able to guarantee its workability on PFA-fixed tissue. Although both other clones i.e. 30-F11 and 5C16, would be good for your samples, CD45 30-F11 # NB100-77417 is a well known clone that offers more flexibility around protocol because of availability of its conjugated forms. It would be advantageous to use a conjugated primary as you are planning for double-immunostaining procedure.

  • Q: For use in Western Blot with CD45 antibodies, what molecular weight of the band should I expect to see?

    A: The theoretical molecular weight for most of our CD45 antibodies is 147 kDa based off the first isoform. Any variation on 147 is due to the immunogen being from a different species and the protein being a slightly different size. CD45 is a family of single chain transmembraneous glycoproteins consisting of at least four isoforms (220, 205, 190, 180 kDa) which share a common large intracellular domain. Their extracellular domains are heavily glycosylated.

  • Q: If this product is used in an application or species as a part of a customer review, will that validate this product in the application/species?

    A: If any of our primary antibodes are used in an untested application or species and it is shown to work through images from customer reviews or through publications, this validates the application/species for this product, allowing the tested application/species to fall under our 100% guarantee. Please check out our Innovator's Reward Program if you decide to test a primary antibody with a species or application that is not currently listed. Please note that the Innovator's Reward Program only applies to our primary antibodies.

  • Q: Is CD45 a good target for use in Neurodegeneration studies?

    A: Yes, here are the approved research areas we have listed for our CD45 products: Adaptive Immunity, Cell Biology, Cellular Markers, Cytokine Research, Glia Markers, Hematopoietic Stem Cell Markers, Immunology, Innate Immunity, Mast Cell Markers, Mesenchymal Stem Cell Markers, Microglia Markers, Myeloid Cell Markers, Myeloid-derived Suppressor, Neurodegeneration, Neuroscience, Signal Transduction, Stem Cell Markers, Growth and Development.

  • Q: We would like to order a CD45 antibody to stain immune cells that were isolated from a ligated sciatic nerve of mice with double immunofluorescence (using PFA-fixed tissue on slides). Which of the following rat monoclonal CD45 antibodies would you recommend: 30-F11, IBL-3/16 or 5C16?

    A:

    I would recommend clone 30-F11. As clone IBL-3/16 has not yet been validated for IHC-P application, we would not be able to guarantee its workability on PFA-fixed tissue. Although both other clones i.e. 30-F11 and 5C16, would be good for your samples, CD45 30-F11 # NB100-77417 is a well known clone that offers more flexibility around protocol because of availability of its conjugated forms. It would be advantageous to use a conjugated primary as you are planning for double-immunostaining procedure.

Showing  1 - 4 of 4 FAQs Showing All
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