Positive control (cell lysate), for antibodies to NALP1 (human)



ALX-840-603-R100	100 µl	49.00 USD
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Product Specification

Alternative Name:	NACHT-, LRR- and PYD-containing protein 1, DEFCAP, NAC, CARD7, Caspase recruitment domain-containing protein 7

Source:	Whole cell lysate of HEK 293T cells transfected with an expression plasmid for human NLRP1/NALP1 (aa 1-1429).

Quantity:	100µl corresponding to 2 x 10⁶ 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 NLRP1/NALP1,mAb (human) (Nalpy1-4) (Prod. No. ALX-804-803), NLRP1/NALP1, pAb (human) (AL176) (Prod. No. ALX-210-904), NLRP1/NALP1, pAb (human) (CT) (Bur 242) (Prod. No. ALX-210-017) and NLRP1/NALP1, pAb (human) (NT) (Bur 241) (Prod. No. ALX-210-018). 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.

Positive control (cell lysate), for antibodies to NALP1 (human) SDS-PAGE

Figure: Detection of full-length human NLRP1/NALP1 (aa 1-1429) with PAb to NLRP1/NALP1 (human) (NT) (Bur 241) (Prod. No. ALX-210-018) 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 NLRP1/NALP1 expression plasmid (Prod. No. ALX-840-603) (lane 2).Method: 13µl of water and 2µl of DTT (100mM) were added to 5µl of cell lysate (diluted 10x Prod. No. ALX-840-603) (corresponding to 10⁵ cells) and boiled 5 min before being loaded on a 8% SDS-PAGE gel. The gel was run at 200V and proteins transferred to nitrocellulose (Amersham Biosciences, Prod. No. 25006565) first for 30 min at 100V in transfer buffer (3g/l trizma-base, 14.4g/l glycin, 20% methanol at pH 8.3) containing 0.1% SDS and then for 45 min. in normal transfer buffer. The nitrocellulose membrane was then blocked in PBS-Tween 0.1%, milk 4% at 4°C and incubated with PAb to NLRP1/NALP1 (human) (NT) (Bur 241) (Prod. No. ALX-210-018) diluted 1:1’000 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 goat anti-rabbit HRP 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 45 sec. with a Fujimedical X-Ray film (Fujifilm, Prod. No. 57164053)."

Figure: Detection of full-length human NLRP1/NALP1 (aa 1-1429) with PAb to NLRP1/NALP1 (human) (CT) (Bur 242) (Prod. No. ALX-210-017) 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 NLRP1/NALP1 expression plasmid (Prod. No. ALX-840-603) (lane 2).Method: 13µl of water and 2µl of DTT (100mM) were added to 5µl of cell lysate (diluted 10x Prod. No. ALX-840-603) (corresponding to 10⁵ cells) and boiled 5 min before being loaded on a 8% SDS-PAGE gel. The gel was run at 200V and proteins transferred to nitrocellulose (Amersham Biosciences, Prod. No. 25006565) first for 30 min at 100V in transfer buffer (3g/l trizma-base, 14.4g/l glycin, 20% methanol at pH 8.3) containing 0.1% SDS and then for 45 min in normal transfer buffer.The nitrocellulose membrane was then blocked in PBS-Tween 0.1%, milk 4% at 4°C and incubated with PAb to NLRP1/NALP1 (human) (CT) (Bur 242) (Prod. No. ALX-210-017) diluted 1:10’000 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 goat anti-rabbit HRP 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 min. with a Fujimedical X-Ray film (Fujifilm, Prod. No. 57164053)."

Figure: Detection of full-length human NLRP1/NALP1 (aa 1-1429) with PAb to NLRP1/NALP1 (human) (AL176) (Prod. No. ALX-210-904) 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 NLRP1/NALP1 expression plasmid (Prod. No. ALX-840-603) (lane 2).Method: 13µl of water and 2µl of DTT (100mM) were added to 5µl of cell lysate (diluted 10x Prod. No. ALX-840-603) (corresponding to 10⁵ cells) and boiled 5 min before being loaded on a 8% SDS-PAGE gel. The gel was run at 200V and proteins transferred to nitrocellulose (Amersham Biosciences, Prod. No. 25006565) first for 30 min at 100V in transfer buffer (3g/l trizma-base, 14.4g/l glycin, 20% methanol at pH 8.3) containing 0.1% SDS and then for 45 min in normal transfer buffer. The nitrocellulose membrane was then blocked in PBS-Tween 0.1%, milk 4% at 4°C and incubated with PAb to NLRP1/NALP1 (human) (AL176) (Prod. No. ALX-210-904) diluted 1:1’000 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 goat anti-rabbit HRP 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 min. with Fujimedical X-Ray film (Fujifilm, Prod. No. 57164053)."

Figure: Detection of full-length human NLRP1/NALP1 (aa 1-1429) with MAb to NLRP1/NALP1 (human) (Nalpy1-4) (Prod. No. ALX-804-803) 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 NLRP1/NALP1 expression plasmid (Prod. No. ALX-840-603) (lane 2).Method: 13µl of water and 2µl of DTT (100mM) were added to 5µl of cell lysate (diluted 10x Prod. No. ALX-840-603) (corresponding to 10⁵ cells) and boiled 5 min before being loaded on a 8% SDS-PAGE gel. The gel was run at 200V and proteins transferred to nitrocellulose (Amersham Biosciences, Prod. No. 25006565) first for 30 min at 100V in transfer buffer (3g/l trizma-base, 14.4g/l glycin, 20% methanol at pH 8.3) containing 0.1% SDS and then for 45 min. in normal transfer buffer. The nitrocellulose membrane was then blocked in PBS-Tween 0.1%, milk 4% at 4°C and incubated with MAb to NLRP1/NALP1 (human) (Nalpy1-4) (Prod. No. ALX-804-803) 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 5 min. with Fujimedical X-Ray film (Fujifilm, Prod. No. 57164053)."

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