discontinued

AM26680FC-N CD165 antibody

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50 µg / €250.00

Quick Overview

Mouse anti Human CD165 AD2

AM26680FC-N

Product Description for CD165

Mouse anti Human CD165 AD2.
Product is tested for Flow Cytometry.

Properties for CD165

Product Category Primary Antibodies
Quantity 50 µg
Synonyms AD2, gp37
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
Add. information This product was originally produced by MBL International.
Application Flow cytometry: 20 L (ready for use); see protocol below.
Background CD44 (H-CAM/Pgp-1/Hermes antigen/ECMR-III/HUTCH-I) is a highly glycosylated transmembrane protein expressed by lymphocytes, fibroblasts, smooth muscle cells, and epithelial cells. CD44 functions as lymphocyte adhesion molecule, acting as a matrix receptor that mediates cell adhesion to the extracellular matrix. CD44 is also involved in T-lymphocyte activation, lymphocyte homing, cell migration, and hemopoiesis. Expression of CD44 on the cell surface changes profoundly during tumor metastasis, and the transition from non-metastatic to metastatic tumor cell variants is associated with expression of CD44 variants (CD44v’s), making CD44 a potential cancer marker.
Concentration 50 µg/ml
Protocols

Flow cytometric analysis for floating cells
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 washing buffer (5x106 cells/mL).

3) Add 50 L of the cell suspension into each tube, and centrifuge at 500 x g for 1 minute at room temperature (20~25oC). Remove supernatant by careful aspiration.
4) Add 10 L of normal goat serum containing 1 mg/mL normal human IgG and 0.1% NaN3 to the cell pellet after tapping. Mix well and incubate for 5 minutes at room temperature.
5) Add 20 L of PE labeled CD44 (15C6). Mix well and incubate for 20 minutes at room temperature.
6) Add 1 mL of the washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration.
7) Resuspend the cells with 500 L of the washing buffer and analyze by a flow cytometer.

Flow cytometric analysis for whole blood cells
We usually use Fisher tubes or equivalents as reaction tubes for all step described below.
1) Add 20 L of PE labeled CD44 monoclonal antibody (15C6) into each tube.
2) Add 100 L of whole blood into each tube. Mix well, and incubate for 30 minutes at room temperature (20~25 oC).
3) Add 1 mL of washing buffer [PBS containing 2% fetal calf serum (FCS) and 0.1% NaN3] followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration.
4) Lyse with OptiLyse C (for analysis on Beckman Coulter instruments) or OptiLyse B (for analysis on BD instruments), using the procedure recommended in the respective package inserts.
5) Add 1 mL of H2O to each tube and incubate for 10 minutes at room temperature.
6) Centrifuge at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration.
7) Add 1 mL of washing buffer followed by centrifugation at 500 x g for 1 minute at room temperature. Remove supernatant by careful aspiration.
8) Resuspend the cells with 500 L of the washing buffer and analyze by a flow cytometer.

 

General Readings
  1. Kozaki, K., et al., Cancer Res. 60, 2535-2540 (2000).
  2. Sugiyama, K., et al., Immunol Invest. 28, 185-200 (1999).
Storage

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

Format
Purification:
Protein-A Sepharose
Buffer System:
PBS
Preservatives:
0.09% NaN3
Stabilizers:
1% BSA
State:
Liquid Ig fraction
Species Reactivity
Species reactivity (tested):
Human
Specificity
Specificity:
This antibody reacts with CD44.

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