Alexa Fluor® 647 (AF6) Conjugated anti-SAM Monoclonal Antibody Clone 118-6 | MAF00201

Alexa Fluor® 647 (AF6) Conjugated anti-SAM Monoclonal Antibody Clone 118-6 | MAF00201 | Arthus Biosystems

Product name        

Alexa Fluor 647-anti-SAM la/b

Catalog Number     

MAF00201-25/50

Description              

Alexa Fluor®647 (AF647) conjugated anti-S-adenosylnnethionine monoclonal antibody clone 118-6

Specificity                

MAF00201 shows the same specificity as un-conjugated mouse anti-SAM monoclonal antibody MA00201.

Properties

Form                                   

Liquid

Storage instructions          

Store at 2-8°C in dark, do not freeze.

Concentration                     

2-4nng/ml,  lot specific

Storage buffer                    

50mM Tris, 150mM NaCI, pH8.0, 0.2% BSA (Sigma), 0.09%NaN3

Dilution buffer                   

PBS, pH 7.4, 1% fetal bovine serum or 0.5% BSA, 0.09%NaN3

Purity                                 

>95% purified with Sephacryl S-200, free from un-conjugated antibody and Alexa Fluor® 647

Clonality                             

Monoclonal

Clone number                     

118-6

Immunoglobin isotype      

mouse IgG2b

Research Areas                  

• Methylation of biomolecules (DNA, RNA, proteins, hormones, neurotransmitters, etc.)
• One-carbon metabolism
• Signal Transduction
• Metabolism
• Pathways and Processes Cancer
• Arthritis
• Neurodegenerative diseases
• Atherosclerosis
• Liver diseases
• Kidney diseases

Applications

The use of MAF00201 in the following applications has been tested. The application notes include recommended and tested dilutions. Optimal dilutions/concentrations should be determined by the end user based on the test environment and purposes.

Recommended

1. Flow Cytometry (FCM):  4-18 pg/nnl
2. Immunofluorescence Laser Scanning Confocal Microscopy (LSCM): 4-8 pg/ml

Figure 1: Immunofluorescence (IF) staining of HepG2 and L02 cells double stained with AF488-anti-SAH 301-3 (Cat# MAF00301) at 40pg/m1 and AF647-anti-SAM 118-6 (Cat#  MAF00201) at 8pg/ml followed by DAPI staining and photographed under the laser scanning confocal microscope (Zeiss LSM 780). (A1-A4) Normal liver cell line L02 cells cultured in RPMI 1640 with 10% FBS for 40h. Different views are as follows: DAPI (A1); AF488 for SAH (A2); AF647 for SAM (A3); Overlap of all the three fluorescent signals  (A4). (B1-B4) Hepatocellular carcinoma cell line HepG2 cells cultured for 40h. Different views are follows: DAPI (B1); AF488 for SAH (B2); AF647 for SAM (B3); Overlap of all the three fluorescent signals (B4). Expression patterns of SAM and SAH are different between L02 and HepG2 cells (x630).

Figure 2: Flow Cytometry of L02 (A1-A3) and HepG2 (B1-B3) cells double stained with Alexa Fluor®488 conjugated anti-SAH antibody 301-3 (Cat# MAF00301) at 36 pg/ml and Alexa Fluor® 647 conjugated anti-SAM antibody 118-6 (Cat# MAF00201) at 4.5 pg/ml. Color legend: Orange:  blank;  Blue:  nuclear fixation/permeabilization  buffer was usedb(eBioscience 00-5523  FoxP3_TF  Staining  Buffer Set);  Red: intracellular fixation/permeabilization buffer was used (eBioscience 00-8824). 100% confluent cells (cultured in RPMI 1640 with 10% FBS for 48h) were fixed and permeabilized with the intracellular fixation/permeabilization buffer (A2, B2) or the nuclear fixation/permeabilization buffer (A3, B3) and then double stained with antibodies indicated above. Cells were used for analysis with BD FACSCalibur Flow Cytometer. SAM expression is higher in L02 than HepG2 cells. Both SAM and SAH are expressed ubiquitously yet rather dynamically. The level of SAM is higher than that of SAH in both cells.