I cannot find any anti-mouse CD45RO for FACS. Have any of your anti-Human CD45RO been tested on mouse T cells?

Unfortunately, none of our CD45RO antibodies for Flow Cytometry have been validated on mouse. If you would be interested in testing the antibody on mouse, we would recommend Our Innovators Reward Program. Under the terms and conditions of this program we would be happy to provide you with a credit of equal or lesser value, in exchange for your online review including an image of either positive or negative staining for testing in a new species and/or application. Please contact us at innovators@novusbio.com with any questions regarding this program.

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?

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.

Human CD45RA Antibody

Name: Human CD45RA Antibody
Brand: R&D Systems
SKU: MAB11446

R&D Systems, part of Bio-Techne | Catalog # MAB11446

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
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Key Product Details

Species Reactivity

Human

Applications

Flow Cytometry, Immunohistochemistry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG₁ Clone # 1069248

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Product Specifications

Immunogen

Synthetic Peptide
Accession # P08575

Specificity

Detects hCD45RA in direct ELISA.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG₁

Scientific Data Images for Human CD45RA Antibody

Detection of CD45RA in PBMC lymphocytes by Flow Cytometry.

PBMC lymphocytes were stained with Mouse Anti-Human CD45RO PE-conjugated Monoclonal Antibody (Catalog # FAB10642P) and either (A) Mouse Anti-Human CD45RA Monoclonal Antibody (Catalog # MAB11446) or (B) Mouse IgG₁ Isotype Control (Catalog # MAB002) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). View our protocol for Staining Membrane-associated Proteins.

Detection of CD45RA in Human Tonsil.

CD45RA was detected in immersion fixed paraffin-embedded sections of Human Tonsil using Mouse Anti-Human CD45RA Monoclonal Antibody (Catalog # MAB11446) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surface in lymphocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Applications for Human CD45RA Antibody

Application

Recommended Usage

Flow Cytometry

0.25 µg/10⁶ cells
Sample: PBMC lymphocytes

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of Human Tonsil

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: CD45RA

CD45RA is a single chain type I glycoprotein that is encoded by the human PTPRC gene. In humans, CD45RA is one of six isoforms of the CD45 complex. These different isoforms are the result of alternative splicing and some of the other isoforms include CD45RO, CD45RB, and CD45RC. CD45RO, CD45RB, CD45RC, and CD45RA are primarily expressed on T cells. CD45RA contains exons 3, 4, and 7 but lacks both exons 5 and 6; CD45RC contains exons 3, 6, and 7. CD45RO contains exons 3 and 7 whereas CD45RB contains exons 3, 5, and 7. Both CD45RA and CD45RB are highly expressed on naïve T cells whereas CD45RO is expressed on activated and memory T cells in addition to some B cell subsets and activated monocytes. Both CD45RA and CD45RO are routinely used to characterize different subsets of Memory T cells.

Long Name

CD45R, A isoform

Alternate Names

B220, CD45 antigen, CD45R, EC 3.1.3.48, L-CA, LY5, protein tyrosine phosphatase, receptor type, C, receptor-type tyrosine-protein phosphatase C, T200 glycoprotein, T200 leukocyte common antigen, T200receptor type, c polypeptide

Entrez Gene IDs

5788 (Human)

Gene Symbol

PTPRC

UniProt

P08575

Additional CD45RA Products

Product Documents for Human CD45RA Antibody

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Product Specific Notices for Human CD45RA Antibody

For research use only

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Flow Cytometry