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

 
ALX-840-607-R100 100 µl 48.00 USD
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Product Specification

Alternative Name:A-proliferating-inducing ligand, TNFSF 13, CD256
 
Source:Whole cell lysate of HEK 293T cells transfected with an expression plasmid for human APRIL (aa 1-250) fused at the C-terminus  to a FLAG®-tag.
 
Quantity:100µl corresponding to 2x105 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 APRIL (human) (Aprily-1) (Prod. No. ALX-804-148), MAb to APRIL (human) (Aprily-5) (Prod. No. ALX-804-801, ALX-804-801B) and MAb to APRIL (human) (Aprily-8) (Prod. No. ALX-804-149).
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.
 
840-607 1
Figure: Detection of full-length human APRIL (aa 1-250) with MAb to APRIL (human) (Aprily-1) (Prod. No. ALX-804-148) 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 APRIL expression plasmid (Prod. No. ALX-840-607) (lane 2).Method: 13µl of water and 2µl of DTT (100mM) were added to 5µl of cell lysate (diluted 10x Prod. No. ALX-840-607) (corresponding to 104 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 1 hour 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 APRIL (human) (Aprily-1) (Prod. No. ALX-804-148) 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 mouse IgG1 (Prod. No. ALX-211-203) 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).
840-607 2
Figure: Detection of full-length human APRIL (aa 1-250) with MAb to APRIL (human) (Aprily-5) (Prod. No. ALX-804-801) 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 APRIL expression plasmid (Prod. No. ALX-840-607) (lane 2).Method: 13µl of water and 2µl of DTT (100mM) were added to 5µl of cell lysate (diluted 10x Prod. No. ALX-840-607) (corresponding to 104 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 1 hour 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 APRIL (human) (Aprily-5) (Prod. No. ALX-804-801) 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 mouse IgG1 (Prod. No. ALX-211-203) 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).
840-607 3
Figure: Detection of full-length human APRIL (aa 1-250) with MAb to APRIL (human) (Aprily-8) (Prod. No. ALX-804-149) 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 APRIL expression plasmid (Prod. No. ALX-840-607) (lane 2).Method: 13µl of water and 2µl of DTT (100mM) were added to 5µl of cell lysate (diluted 10x Prod. No. ALX-840-607) (corresponding to 104 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 1 hour 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 APRIL (human) (Aprily-8) (Prod. No. ALX-804-149) 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 mouse IgG1 (Prod. No. ALX-211-203) 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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