Thermo Fisher Scientific SEMA4A Monoclonal Antibody (5E3), PerCP-eFluor 710, eBioscience

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

5 µL (0.06 µg)/test

-

Product Specifications

Species Reactivity

Human, Mouse

Host/Isotype

Mouse / IgG1, kappa

Recommended Isotype Control

Mouse IgG1 kappa Isotype Control (P3.6.2.8.1), PerCP-eFluor™ 710, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

5E3

Conjugate

PerCP-eFluor™ 710 PerCP-eFluor™ 710 PerCP-eFluor™ 710

Excitation/Emission Max

482/708 nm View spectra

Form

Liquid

Concentration

5 µL/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_2573898

Product Specific Information

Description: The monoclonal antibody 5E3 reacts with human and mouse Semaphorin 4A (Sema4A). This 150 kDa glycoprotein is a member of the class IV semaphorin family and exists as membrane-bound and soluble forms. Sema4A is expressed by a wide variety of tissues, including lung, kidney, spleen, testis, and mammary glands, as well as activated T cells and dendritic cells. This ligand binds to Tim-2, neuropilin-1 (Nrp1), plexin D1, and plexin B1. Sema4A plays a role in T cell activation, Th1 differentiation, and Treg function and survival.

Our testing demonstrates that the 5E3 antibody stains resting CD49b+ NK cells in mouse. However, staining of CD56+ NK cells in human is negative.

Applications Reported: This 5E3 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This 5E3 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.06 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.

PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.

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: 488 nm; Emission: 710 nm; Laser: Blue Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

SEMA4A is a member of the semaphorin family of soluble and transmembrane proteins. Semaphorins are involved in guidance of axonal migration during neuronal development and in immune responses.

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