Invitrogen

CD45R (B220) Monoclonal Antibody (RA3-6B2), Super Bright™ 780, eBioscience™

Product Image Gallery

CD45R (B220) Antibody in Flow Cytometry (Flow) — Staining of C57BL/6 splenocytes with CD3e Monoclonal Antibody, FITC (Product # 11-0031-82) and 0.25 µg of Rat IgG2a kappa Isotype Control, Super Bright 780 (Product # 78-4321-82) (left) or 0.25 µg of CD45R (B220) Monoclonal Antibody, Super Bright 780 (right). Total viable cells were used for analysis.

CD45R (B220) Antibody in Immunohistochemistry, Immunohistochemistry (Paraffin) (IHC, IHC (P)) — Figure 5 Innate and adaptive immune cell responses in the colon following PACAP treatment of C. jejuni infected secondary abiotic IL-10 -/- mice. Mice were perorally infected with C. jejuni strain 81-176 by gavage on day (d) 0 and d1, and subjected to intraperitoneal treatment with either synthetic PACAP or vehicle (mock) from d2 until d5 post-infection. On d6 the average numbers of ( A ) macrophages and monocytes (F4/80 + ), ( B ) T lymphocytes (CD3 + ), ( C ) regulatory T cells (FOXP3 + ) and ( D ) B lymphocytes (B220 + ) were quantitated in six high power fields (HPF) of colonic paraffin sections applying immunohistochemistry. Naive mice were used as negative controls. Box plots indicate the 75th and 25th percentiles of the median (black bar within boxes). The total range, the significance levels ( p values calculated by the one-way ANOVA test followed by the Tukey post-correction test for multiple comparisons) and the total numbers of mice under investigation (in parentheses) are indicated. Results pooled from four independent experiments are shown.

CD45R (B220) Antibody in Immunohistochemistry (IHC) — Figure 8 Establishment of adaptive immunity in the infected lungs of WT and CXCL5 -/- mice. (A) Lung lobe sections from WT and CXCL5 -/- mice were stained at 10 d.p.i. with an anti-B220 antibody and hematoxylin (x30 magnification). (B) The positively stained area (iBALT structure) in each lung lobe section was calculated using CaseViewer software (3DHISTECH). The total area of iBALT per lung section was calculated from 4 mice per group (3 lung lobe sections per mouse). (C) Lung sections were stained with anti-B220 (red) and anti-CD4 (yellow) antibodies for immunofluorescence detection. The immunofluorescence sections were counterstained with DAPI (blue). (D) GO functional analysis of upregulated genes in the infected lungs of CXCL5 -/- mice compared to WT mice at 7 and 10 d.p.i. The methods and criteria for filtration of differentially expressed genes are described in the MATERIALS AND METHODS section. (E) Mice were infected with H1N1 or H3N2 (3000 CCID 50 ), and the body weight loss (n=12) of the mice was monitored until 2 weeks post infection. WT-1 H1N1: WT mice infected with H1N1; WT-1 H3N2: WT mice infected with H3N2; CXCL5 -/- 1 H1N1: CXCL5 -/- mice infected with H1N1; CXCL5 -/- 1 H3N2: CXCL5 -/- mice infected with H3N2; WT-2 H1N1: WT mice recovered from H1N1 infection and infected with H1N1; WT-2 H3N2: WT mice recovered from H1N1 infection and infected with H3N2; CXCL5 -/- 2 H1N1: CXCL5 -/- mice recovered from H1N1 infection and infected with H1N1; CXCL5 -/- 2 H3N2: CXC

Product Details

78-0452-82

Applications	Tested Dilution	Publications
Immunohistochemistry (IHC)	-	View 14 publications 14 publications
Immunohistochemistry (Paraffin) (IHC (P))	-	View 1 publication 1 publication
Flow Cytometry (Flow)	0.5 µg/test	View 13 publications 13 publications
Miscellaneous PubMed (Misc)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Mouse
Published species	Not Applicable
Host/Isotype	Rat / IgG2a, kappa
Recommended Isotype Control	Rat IgG2a kappa Isotype Control (eBR2a), Super Bright™ 780, eBioscience™
Class	Monoclonal
Type	Antibody
Clone	RA3-6B2
Conjugate	Super Bright™ 780 Alexa Fluor 488 Alexa Fluor 532 Alexa Fluor 561 Alexa Fluor 660 Alexa Fluor 700 APC APC-eFluor 780 Biotin Brilliant UV 395 Brilliant UV 496 Brilliant UV 615 Brilliant UV 661 Brilliant UV 737 Brilliant UV 805 Brilliant Violet 650 Brilliant Violet 711 Brilliant Violet 786 eFluor 450 eFluor 506 eFluor 570 eFluor 615 eFluor 660 FITC Functional Grade NovaFluor Blue 510
Excitation/Emission Max	413/780 nm View spectra
Form	Liquid
Concentration	0.2 mg/mL
Purification	Affinity chromatography
Storage buffer	PBS, pH 7.2, with BSA
Contains	0.09% sodium azide
Storage conditions	4° C, store in dark, DO NOT FREEZE!
RRID	AB_2722971

Product Specific Information

Description: The RA3-6B2 monoclonal antibody reacts with exon A-restricted isoform of mouse CD45, a 220 kDa surface molecule. CD45R/B220 epitope is mainly expressed by the B cell lineage from early Pro-B to mature B cells. However, some activated T cells, lymphokine activated killer cells (LAK), NK cell progenitors in the bone marrow, and T cells of the lpr/lpr mutant mouse also express this antigen.

Applications Reported: This RA3-6B2 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This RA3-6B2 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.5 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Super Bright 780 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 780 nm. We recommend using a 780/60 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome.

When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information.

In some experiments, we have observed that compensation values for Super Bright 780-conjugated antibodies are higher in the violet 450/50 channel when using UltraComp eBeads microspheres (Product # 01-2222-42) as compared to single-color stained cells. In such circumstances, we would recommend setting compensation with cells. We have also observed this in some experiments using AbC Total Antibody Compensation beads (Product # A10497).

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 405 nm; Emission: 780 nm; Laser: Violet Laser

Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.

Target Information

CD45R (PTPRC) is a member of the protein tyrosine phosphatase (PTP) family. PTPs are known to be signaling molecules that regulate a variety of cellular processes including cell growth, differentiation, mitotic cycle, and oncogenic transformation. CD45 contains an extracellular domain, a single transmembrane segment and two tandem intracytoplasmic catalytic domains. CD45 is specifically expressed in hematopoietic cells and has been shown to be an essential regulator of T- and B-cell antigen receptor signaling. CD45 glycoprotein is crucial in lymphocyte development and antigen signaling, serving as an important regulator of Src-family kinases. CD45 protein exists in multiple isoforms as a result of alternative splicing, and these isoforms differ in their extracellular domains, but share identical transmembrane and cytoplasmic domains. Further, these CD45 isoforms differ in their ability to translocate into the glycosphingolipid-enriched membrane domains and their expression depends on cell type and physiological state of the cell. In addition to its role in immunoreceptor signaling, CD45 is important in promoting cell survival by modulating integrin-mediated signal transduction pathway and is also involved in DNA fragmentation during apoptosis. CD45RA is an isoform of the CD45 complex and has restricted expression between different subtypes of lymphoid cells. Diseases associated with CD45 dysfunction include Severe Combined Immunodeficiency (T-cell negative, B-Cell Negative) and Hepatitis C.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Flow Cytometry

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Bioinformatics

Protein Aliases: CD45; L-CA; Leukocyte common antigen; Ly-5; Lymphocyte antigen 5; lymphocyte common antigen; Receptor-type tyrosine-protein phosphatase C; T200

Gene Aliases: B220; Cd45; CD45R; L-CA; loc; Ly-5; Lyt-4; Ptprc; T200

UniProt ID: (Mouse) P06800

Entrez Gene ID: (Mouse) 19264

Molecular Function: protein phosphatase protein modifying enzyme

Function(s)

Process(es)

phosphoprotein phosphatase activity protein tyrosine phosphatase activity protein binding heparin binding hydrolase activity phosphatase activity protein kinase binding heparan sulfate proteoglycan binding

CD45R (B220) Monoclonal Antibody (RA3-6B2), Super Bright™ 780, eBioscience™

Product Details

78-0452-82

Immunohistochemistry (IHC)

Immunohistochemistry (Paraffin) (IHC (P))

Flow Cytometry (Flow)

Miscellaneous PubMed (Misc)

Species Reactivity

Published species

Host/Isotype

Recommended Isotype Control

Class

Type

Clone

Conjugate

Excitation/Emission Max

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

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