CXCR4 (extracell. Dom.) (N-term) Rat Monoclonal Antibody [Clone ID: A145]

CAT#: AM26473AF-N

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CXCR4 (extracell. Dom.) (N-term) rat monoclonal antibody, clone A145, Azide Free



Size
    • 100 ug


Product images

Specifications

Product Data
Clone Name A145
Applications FC
Recommended Dilution Flow cytometry: 10-20 μg/ml (final concentration). For details see protocols below.
Reactivity Human
Host Rat
Isotype IgG1
Clonality Monoclonal
Immunogen Cos-1 cells transfected with CXCR4 gene
Specificity

The antibody recognizes the N-terminal, extracellular domain of the CXCR4 protein.

Formulation PBS containing 50% glycerol. Contains no preservatives.
State: Azide Free
State: Liquid Ig fraction
Concentration lot specific
Purification Ammonium sulfate precipitation followed by gel filtration through Superdex 200 in PBS
Storage

Store undiluted at 2-8°C for one month or (in aliquots) at -20°C for longer.
Avoid repeated freezing and thawing.

Stability Shelf life: one year from despatch.

Synonyms CXC-R4, CXCR-4, Fusin, LCR1, FB22, NPYRL, HM89, SDF1 receptor, LESTR
Note This product was originally produced by MBL International.

Protocol:

Flow cytometric analysis for floating cells

Protocol 1

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 (5x10e6 cells/mL).
3) Add 50 µL of the cell suspension into each tube, and centrifuge at 500xg for 1 minute at room temperature (20~25oC). Remove supernatant by careful aspiration.
4) Add 10 µL 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 (20~25 oC).
5) Add 30 µL of the anti-CXCR4 monoclonal antibody (10-20 µg/mL) diluted with the washing buffer. Mix well, and incubate for 30 minutes at room temperature (20~25 oC).
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) Add 30 µL of secondary antibody (1:40 FITC conjugated anti-Rat IgG) diluted with the washing buffer. 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 µL of the washing buffer and analyze by a flow cytometer.

Protocol 2

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, 1 mg/mL normal human IgG and 0.1% NaN3 (5x106 cells/mL).
3) Add 20 µL of the anti-CXCR4 monoclonal antibody (50 µg/mL) diluted with the washing buffer into each tube.
4) Add 50 µL 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 for 1 minute at room temperature (20~25oC). Remove supernatant by careful aspiration.
6) Resuspend the cells with 50 µL of the washing buffer.
7) Add 20 µL of secondary antibody (1:10 FITC conjugated anti-Rat 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 µL of the washing buffer and analyze by a flow cytometer.

 






 

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*Delivery time may vary from web posted schedule. Occasional delays may occur due to unforeseen complexities in the preparation of your product. International customers may expect an additional 1-2 weeks in shipping.
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