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Test sample

Test sample in the group Alla produktegenskaper at Agrisera AB (Antibodies for research) (AS20 Sample)



DATA SHEET IN PDF

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How to cite this product:
Product name, number (Agrisera, Sweden)

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Reactivity

Application examples

Application examples

Thank you for participating in Agrisera Antibody Testing Program!

Recommended protocol:

  • 20 ug protein load/well
  • Blocking: 5 % non-fat milk 1h/RT
  • Primary antibodies: 1: 1000 ON/4C TBS-T
  • Secondary antibodies (Agrisera): 1: 25 000 1h/RT
  • Detection: AgriseraSuperBright (for proteins of low expression) or AgriseraBright (for proteins of moderate expression)

 

Results are to be returned in the following format:

Description of each lane (as in the example below):

1 - 50 ug of Arabidopsis thaliana whole leaf extract

2 - 50 ug of Arabidopsis thaliana mutant

Mark: MW markers: in example below: MagicMark from Invitrogen

 

Mark MW of a target protein to be detected

 

Provide an IMAGE (whole membrane with MW markers), of a good quality, like in the example below:

 

 

Fill in ALL experimental details in the template below:

µg/well of total protein extracted freshly from    with write exact buffer components ..... and denatured with....at °C or 70°C for 2-5 min.... were separated on …. % SDS-PAGE  and blotted 1h to PVDF/nitrocellulose (pore size of    um), using wet, semi-dry or dry transfer. Blot was blocked with % milk or % BSA  for 1h/RT or 4°C/ON with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h/RT with agitation in TBS-T or ON/4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:25 000 in  for 1h/RT with agitation. The blot was washed as above and developed for   min  with Agrisera ECLBright or AgriseraECLSuperBright with   name and supplier. Exposure time was.....seconds.

 

Questions to answer:

Was used material freshly extracted or stored?

Was a blocking reagent prepared freshly?

How was primary antibody stored?

Was primary antibody or any other reagent re-used?

Was incubation volume for each step kept the same?

 

Thank you for your contribution!

In return we offer a 10 % discount on your next order from Agrisera

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