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



ALX-840-605-R100	100 µl	48.00 USD

Product Specification

Alternative Name:	BlyS, THANK, TALL-1, zTNF4, TNFSF 13B/20, CD257

Quantity:	100µl corresponding to 2 x 10⁵ 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).

Source/Host:	Whole cell lysate of HEK 293T cells transfected with an expression plasmid for human BAFF (aa 83-285).

Application:	Positive control in Western blot for MAb to BAFF (human) (Buffy-1) (Prod. No. ALX-804-128, ALX-804-128F) and MAb to BAFF (Buffy-2) (Prod. No. ALX-804-131, ALX-804-131F). Add 13µl sterile water and 2µl DTT (100mM) to 5µl of cell lysate.

Long Term Storage:	-20°C

Handling:	After opening, prepare aliquots and store at -20°C. Avoid freeze/thaw cycles.

Figure: Detection of full-length human BAFF (aa 83-285) with MAb to BAFF (human) (Buffy-1) (Prod. No. ALX-804-128) 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 BAFF expression plasmid (Prod. No. ALX-840-605) (lane 2).Method: 13µl of water and 2µl of DTT (100mM) were added to 5µl of cell lysate (diluted 10x Prod. No. ALX-840-605) (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 BAFF (human) (Buffy-1) (Prod. No. ALX-804-128) 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 5 sec. with Fujimedical X-Ray film (Fujifilm, Prod. No. 57164053).

Figure: Detection of full-length human BAFF (aa 83-285) with MAb to BAFF (Buffy-2) (Prod. No. ALX-804-131) 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 BAFF expression plasmid (Prod. No. ALX-840-605) (lane 2).Method: 13µl of water and 2µl of DTT (100mM) were added to 5µl of cell lysate (diluted 10x Prod. No. ALX-840-605) (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 BAFF (Buffy-2) (Prod. No. ALX-804-131) 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 1 min. with Fujimedical X-Ray film (Fujifilm, Prod. No. 57164053).

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