SIGLEC5 antibody (FITC)

Catalog No. ABIN5541400

This anti-SIGLEC5 antibody is a Rat Monoclonal antibody detecting SIGLEC5 in FACS. Suitable for Mouse.

Quick Overview for SIGLEC5 antibody (FITC) (ABIN5541400)

Target: SIGLEC5 (Sialic Acid Binding Ig-Like Lectin 5 (SIGLEC5))

Reactivity: Mouse

Host: Rat

Clonality: Monoclonal

Conjugate: This SIGLEC5 antibody is conjugated to FITC

Application: Flow Cytometry (FACS)

Clone: 8D2

Product Details anti-SIGLEC5 Antibody (FITC)

Specificity: This antibody reacts with mouse SiglecE.

Purification: Protein G agarose

Immunogen: Mouse SiglecE transfected L cell

Isotype: IgG2b

Application Details

Application Notes: Flow cytometry: 50 μg/mL (final concentration). For details see protocol below.

Protocol: Flow cytometric analysis for floating cells Protocol 1 We usually use Fisher tubes or equivalents as reaction tubes for all step described below. 1) Wash the cells 3 times with washing buffer [PBS containing 2 % fetal calf serum (FCS) and 0.1 % NaN 3]. 2) Resuspend the cells with washing buffer (5x10 6 cells/mL). 3) Add 50 μ L of the cell suspension into each tube, and centrifuge at 500 x g for 1 minute at room temperature (20~25 o C). Remove supernatant by careful aspiration. 4) Add 10 μ L of normal goat serum containing 1 mg/mL normal human IgG and 0.1 % NaN 3 to the cell pellet after tapping. Mix well and incubate for 5 minutes at room temperature (20~25 o C). 5) Add 20 μL of the FITC labeled anti-mouse SiglecE monoclonal antibody (8D2) (50 μ g/mL) diluted with the washing buffer. Mix well and incubate for 30 minutes at room temperature (20~25 o C). 6) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature (20~25 o C). Remove supernatant by careful aspiration. 7) Add 20 μL of 1:50 PE conjugated anti-mouse CD11c (Pharmingen: code no.557401) diluted with the washing buffer. Mix well and incubate for 15 minutes at room temperature (20~25 o C). 8) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature (20~25 o C). Remove supernatant by careful aspiration. 9) Resuspend the cells with 500 μ L of the washing buffer and analyze by a flow cytometer. Positive control for flow cytometry mouse splenocytes Protocol 2 We usually use Fisher tubes or equivalents as reaction tubes for all steps described below. 2) Resuspend the cells with PBS containing 25 % normal goat serum, 1 mg/mL normal human IgG and 0.1 % NaN 3 (5x10 6 cells/mL). 3) Add 20 μ L of the FITC labeled anti-mouse SiglecE monoclonal antibody (8D2) (50 μ g/mL) diluted with the washing buffer into each tube. 4) Add 50 μ L of the cell suspension into each tube. Mix well and incubate for 30 minutes at room temperature (20~25 o C). 5) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature (20~25 o C). Remove supernatant by careful aspiration. 6) Resuspend the cells with 50 μ L of the washing buffer.

Restrictions: For Research Use only

Handling

Format: Liquid

Storage: 4 °C

Storage Comment: Store at 2-8 °C. Shelf life: one year from despatch.

Expiry Date: 12 months

Target Details for SIGLEC5

Target: SIGLEC5 (Sialic Acid Binding Ig-Like Lectin 5 (SIGLEC5))

Alternative Name: cd170,siglec5

Background: SilecE (sialic acid-binding immunoglobulin-like lectin E/SigLec5/CD170) is a putative adhesion molecule that mediates sialic-acid dependent binding to cells. SiglecE is expressed on neutrophil and monocyte popul ations, both in the blood and bone marrow, as a dimeric, disulfide linked, 140 kDa type I membrane protein containing cytoplasmic immune receptor-based tyrosi ne signaling motifs. SiglecE recruits the SH2-domain-containing protein tyrosine phosphatases SHP-1 and SHP-2, which block signal transduction through dephosphorylation of signaling molecules. Thus SiglecE acts as an inhibitory receptor for ligand induced tyrosine phosphorylation.

UniProt: Q920G3