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Positive control (cell lysate), for antibodies to RIP2 (human)

 
ALX-840-604-R100 100 µl 49.00 USD
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Product Specification

Alternative Name:CARDIAK, CARD-containing IL-1β converting enzyme associated kinase, RICK, RIPK2, Receptor-interacting serine-threonine kinase 2, Receptor-interacting protein 2
 
Source:Whole cell lysate of HEK 293T cells transfected with an expression plasmid for human RIP2 (receptor-interacting protein 2) (aa 1-541) fused at the C-terminus to a FLAG®-tag.
 
Quantity:100µl corresponding to 2x106 cells.
 
Formulation:Liquid. In 5x sample buffer (312.5mM TRIS, pH 6.8, containing 10% SDS, 6% glycerol, 0.075% bromophenol blue, 10% β-mercaptoethanol and 10mM DTT).
 
Application Notes:Positive control in Western blot for MAb to RIP2 (human) (Nick-1) (Prod. No. ALX-804-139). Add 13µl sterile water and 2µl DTT (100mM) to 5µl of cell lysate.
 
Shipping:Shipped on Blue Ice
 
Long Term Storage:-20°C
 
Handling:Avoid freeze/thaw cycles. After opening, prepare aliquots and store at -20°C.
 
Technical Info/Product Notes:FLAG is a registered trademark of Sigma-Aldrich Co.
 
840-604
Figure: Detection of full-length human RIP2 (aa 1-541) with MAb to RIP2 (human) (Nick-1) (Prod. No. ALX-804-139) in a cell lysate control of mock transfected cells (HEK 293T) (Prod. No. ALX-840-600) (lane 1) and HEK 293T cells transfected with a full-length human RIP2 expression plasmid (Prod. No. ALX-840-604) (lane 2).Method: 13µl of water and 2µl of DTT (100mM) were added to 5µl of cell lysate (Prod. No. ALX-840-604) (corresponding to 105 cells) and boiled 5 min before being loaded on a 12% SDS-PAGE gel. The gel was run at 200V and proteins transferred to nitrocellulose (Amersham Biosciences, Prod. No. 25006565) for 1hour at 100V in transfer buffer (3g/l trizma-base, 14.4g/l glycin, 20% methanol at pH 8.3).The nitrocellulose membrane was then blocked in PBS-Tween 0.1%, milk 4% at 4°C and incubated with MAb to RIP2 (human) (Nick-1) (Prod. No. ALX-804-139) at 1µg/ml in PBS-Tween 0.1%, milk 4% for 1 hour at RT. After three washes in PBS-Tween 0.1%, the membrane was incubated with PAb to rat IgG (Prod. No. ALX-211-052) diluted 1:5’000 in PBS-Tween 0.1%, milk 4% for 45 min at RT. After three washes in PBS-Tween 0.1%, the membrane was revealed in ECL solution (Pierce, Prod. No. 34080). The membrane was exposed for 30 sec. with Fujimedical X-Ray film (Fujifilm, Prod. No. 57164053).
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840-604

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RIP2 (human) monoclonal antibody (Nick-1) 

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ALX-804-139-C100 100 µg 356.00 USD
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