CD45RA Antibody (Leukocyte marker)

SKU: orb248519

Description

This antibody recognizes a protein of 205kDa-220kDa, identified as CD45RA, an isoform of the human leukocyte common antigen (CD45). Human CD45 contains three exons which encode peptide segments designated A, B and C. The differential splicing of the exons generates at least five isoforms, ABC, AB, BC, B and O. This antibody reacts with ABC and BC isoforms. CD45RA is expressed on 40-50% of peripheral CD4+ Tcells, 50% of peripheral CD8+ T-cells, B-cells, and leukemic B-cell lines. T-cells expressing CD45RA are naive or virgin Tcells. T-cells expressing CD45RO are memory Tcells. CD45RA and CD45RO define complementary, predominantly non-overlapping populations of resting peripheral Tcells. This CD45RA antibody is useful in the study of the subpopulation of CD4+ or CD8+ T-cells. It is especially useful in differentiating T-cell lymphomas (CD45RO +ve) from B cell lymphomas (CD45RA +ve).

Images & Validation

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Tested Applications	FACS, IF, IHC-P, WB
Dilution range	Flow cytometry: 0.5-1ug/million cells,Immunofluorescence: 1-2ug/ml,Western blot: 1-2ug/ml,Immunohistochemistry (FFPE): 0.5-1ug/ml for 30 min at RT
Reactivity	Human
Application Notes	The concentration stated for each application is a general starting point. Variations in protocols, secondaries and substrates may require the CD45RA antibody to be titered up or down for optimal performance.1. The prediluted format is supplied in a dropper bottle and is optimized for use in IHC. After epitope retrieval step (if required), drip mAb solution onto the tissue section and incubate at RT for 30 min.

Key Properties

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Antibody Type	Primary Antibody
Host	Mouse
Clonality	Monoclonal
Isotype	Mouse IgG2a, kappa
Clone No.	158-4D3
Immunogen	Stimulated human leukocytes were used as the immunogen for this CD45RA antibody.
Conjugation	Unconjugated

Storage & Handling

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Storage	Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
Disclaimer	For research use only

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Immunofluorescent staining of PFA-fixed human Raji cells with CD45RA antibody (clone 158-4D3, green) and Reddot nuclear stain (red).

IHC staining of FFPE human tonsil with CD45RA antibody (clone 158-4D3). HIER: steam section in pH6 citrate buffer for 20 min and allow to cool prior to staining.

FACS surface staining of human PBMCs using CD45RA antibody (clone 158-4D3, red), and isotype control (green).

Flow cytometry testing of human Jurkat cells with CD45RA antibody (clone 158-4D3); Red = isotype control, Blue = CD45RA antibody.

Western blot testing of human spleen lysate with CD45RA antibody (clone 158-4D3). Expected molecular weight: 147-220 kDa depending on glycosylation level.

SDS-PAGE analysis of purified, BSA-free CD45RA antibody (clone 158-4D3) as confirmation of integrity and purity.

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Protocol Information

Western Blot (IB, immunoblot)

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IHC-P

Immunohistochemistry Paraffin

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FACS

Fluorescence-Activated Cell Sorting (FC, Flow cytometry)

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Immunofluorescence

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