discontinued

AM26680AF-N CD165 antibody

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0.1 mg / €250.00

Quick Overview

Mouse anti Human CD165 AD2

AM26680AF-N

Product Description for CD165

Mouse anti Human CD165 AD2.
Presentation: Azide Free
Product is tested for Flow Cytometry.

Properties for CD165

Product Category Primary Antibodies
Quantity 0.1 mg
Synonyms AD2, gp37
Presentation Azide Free
Reactivity Hu
Applications F
Clonality Monoclonal
Clone AD2
Host Mouse
Isotype IgG1
Shipping to Not USA/Canada
PDF datasheet View Datasheet
Manufacturer Acris Antibodies GmbH
Material safety datasheet MSDS for Monoclonal Antibodies (de)

Datasheet Extract

Immunogen
Immunogen:
HSB cells
Isotype control AM33027AF-N
Add. information

This product was originally produced by MBL International.

Application Flow cytometry: 10-20 mg/ml (final concentration).
For deteils see protocols below.
Background CD165 or gp37 is a cell surface molecule present on a subset of peripheral lymphocytes and monocytes and is important for adhesion of thymocytes to thymic epithelial cells.
Protocols

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% NaN3].
2) Add 10 mL of normal goat serum to the cell pellet after tapping. Mix well, and incubate for 5 minutes at room temperature (20~25 oC).
3) Add 30 µL of the CD165 monoclonal antibody (10-20 mg/mL) diluted with the washing buffer. Mix well, and incubate for 30 minutes at room temperature (20~25 oC).
4) Add 1 mL of the washing buffer followed by centrifugation at 500xg for 1 minute at room temperature (20~25oC). Remove supernatant by careful aspiration.
5) Add 30 µL of secondary antibody (1:40 FITC conjugated anti-mouse IgG) diluted with the washing buffer. Mix well and incubate for 15 minutes at room temperature (20~25oC).
6) Add 1 mL of the washing buffer followed by centrifugation at 500xg for 1 minute at room temperature (20~25oC). Remove supernatant by careful aspiration.
7) Resespend the cells with 500 mL of the washing buffer and analyze by a flow cytometer.

Protocol 2
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% NaN3].
2) Resuspend the cells with PBS containing 25% normal goat serum and 0.1% NaN3 (5x10e6 cells/ml).
3) Add 20 mL of the CD165 monoclonal antibody (50 mg/mL) diluted with the washing buffer into each tube.
4) Add 50 mL of the cell suspension into each tube. Mix well, and incubate for 30 minutes at room temperature (20~25 oC).
5) Add 1 mL of the washing buffer followed by centrifugation at 500xg rpm for 1 minute at room temperature (20~25oC). Remove supernatant by careful aspiration.
6) Resuspend the cells with 50 mL of the washing buffer.
7) Add 20 µL of secondary antibody (1:10 FITC conjugated anti-mouse IgG) diluted with the washing buffer into each tube. Mix well and incubate for 15 minutes at room temperature (20~25oC).
8) Add 1 mL of the washing buffer followed by centrifugation at 500xg for 1 minute at room temperature (20~25oC). Remove supernatant by careful aspiration.
9) Resuspend the cells with 500 mL of the washing buffer and analyze by a flow cytometer.

General Readings
  1. Bruggers, C.S., et al. J. Immunol. 154, 2012-2022 (1995).
Storage

Store (in aliquots) at -20 °C. Avoid repeated freezing and thawing.
Shelf life: one year from despatch.

Format
Purification:
Protein-A Sepharose
Buffer System:
PBS containing 50% glycerol, without preservatives
State:
Liquid Ig fraction
Azide Free
Species Reactivity
Species reactivity (tested):
Human
Specificity
Specificity:

This antibody reacts with AD2 antigen.

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