Alexa Fluor® 488 (AF488) Conjugated anti-SAH Monoclonal Antibody Clone 301-3 | MAF00301

Alexa Fluor® 488 (AF488) Conjugated anti-SAH Monoclonal Antibody Clone 301-3 | MAF00301 | Arthus Biosystems

Product name        

Alexa Fluor 488-anti-SAH la/b

Catalog Number     

MAF00301-25/50

Description              

Alexa Fluor® 488 (AF488) conjugated anti-S-adenosylhomocysteine monoclonal antibody clone 301-3

Specificity                

MAF00301 shows the same specificity as un-conjugated mouse anti-SAH monoclonal antibody MA00303.

Properties

Form                                   

Liquid

Storage instructions          

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

Concentration                     

2-4ring/m1 or 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 Sephadex G-25, free from un-conjugated antibody and Alexa Fluor® 488

Clonality                             

Monoclonal

Clone number                    

301-3

Immunoglobin isotype      

mouse IgG3

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 MAF00301 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

Flow Cytometry (FCM): 25-80 pg/ml

Immunofluorescence Laser Scanning Confocal Microscopy (LSCM): 20-40 pg/ml

Figure 1: Immunofluorescence (IF) staining of HepG2 and L02 cells double stained with AF488-anti-SAH 301-3 (Cat# MAF00301) at 20pg/m1 and AF647-anti-SAM 118-6 (Cat# MAF00201) at 4pg/ml followed by DAPI staining and photographed under the laser scanning confocal microscope Zeiss LSM 78 (x630). (Al-A4) Normal liver cell line L02 cells cultured in RPM! 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 (100% confluent). Different views are follows: DAPI (B1); AF488 for SAH (B2); AF647 for SAM (B3); Overlap of all the three fluorescent signals (B4). Both antibodies were used at relatively low concentration. When cells are 100% confluent, both SAM and SAH are mainly seen around nuclear membrane.

Figure 2: Flow Cytometry of L02 (Al-A4) and HepG2 (B1-B4) cells double stained with Alexa Fluor® 488 conjugated anti-SAH antibody 301-3 (Cat# MAF00301) at 28 pg/ml and Alexa Fluor® 647 conjugated anti-SAM antibody 118-6 (Cat# MAF00201) at 4.5 pg/ml. 100% confluent cells (cultured for 48h) were fixed and permeabilized with the intracellular fixation/permeabilization buffer (A2, B2) or the nuclear fixation/permeabilization buffer (A4, B4) and then double stained with antibodies indicated above. Al and B1 are the unstained control for A2 and B2 respectively. A3 and B3 are the unstained control for A4 and B4 respectively. Cells were used for analysis with BD FACSCalibur Flow Cytometer. Both fluorescent signals in A4 and B4 are higher than those of A2 and B2.