K2001

Apoptosis Antibody Sampler Kit

cat.: K2001

Product Type:	Antibody Sampler Kit
Species reactivity:	Human
Applications:	WB

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Uniprot #:	SwissProt: P42574 Human \| P42574 Human \| P09874 Human \| P11103 Mouse \| P27008 Rat \| P09874 Human \| P55211 Human \| Q8C3Q9 Mouse \| P55211 Human \| P55210 Human \| P55210 Human
Alternative names:	APAF-3 APAF3 Apoptosis related cysteine peptidase Apoptotic protease Mch-6 Apoptotic protease-activating factor 3 CASP-9 CASP9 CASP9_HUMAN Caspase 9 apoptosis related cysteine peptidase Caspase 9 Dominant Negative Caspase 9c Caspase-9 Caspase-9 subunit p10 ICE LAP6 ICE like apoptotic protease 6 ICE-LAP6 ICE-like apoptotic protease 6 MCH6 PPP1R56 protein phosphatase 1 regulatory subunit 56 RNCASP9

Images

Fig1: Western blot analysis of Caspase-3 on different lysates with Rabbit anti-Caspase-3 antibody (ET1602-39) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 1μM staurosporine for 3 hours cell lysate
Lane 3: Jurkat cell lysate
Lane 4: Jurkat treated with 25μM Etoposide for 5 hours cell lysate
Lane 5: MCF7 cell lysate (negative)
Lane 6: HEK-293 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 32 kDa
Observed band size: 32 kDa

Exposure time: 3 minutes 20 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1602-39) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of PARP on different lysates with Rabbit anti-PARP antibody (ET1608-56) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution.

Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: Jurkat cell lysate (15 µg/Lane)
Lane 3: NIH/3T3 cell lysate (15 µg/Lane)
Lane 4: C2C12 cell lysate (15 µg/Lane)
Lane 5: C6 cell lysate (15 µg/Lane)
Lane 6: PC-12 cell lysate (15 µg/Lane)

Predicted band size: 113 kDa
Observed band size: 113 kDa

Exposure time: 6 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1608-56) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

Fig3: Western blot analysis of Cleaved PARP on different lysates with Rabbit anti-Cleaved PARP antibody (ET1608-10) at 1/2,000 dilution.

Lane 1: HeLa whole cell lysate
Lane 2: HeLa treated with 1μM staurosporine for 3 hours whole cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 89 kDa
Observed band size: 89 kDa

Exposure time: 1 minute 9 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1608-10) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

Fig4: Western blot analysis of Caspase-9 on different lysates with Rabbit anti-Caspase-9 antibody (ET1610-95) at 1/1,000 dilution.

Lane 1: Hela-si NT cell lysate
Lane 2: Hela-si Caspase-9 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 46 kDa
Observed band size: 46 kDa

Exposure time: 2 minutes;

4-20% SDS-PAGE gel.

ET1610-95 was shown to specifically react with Caspase-9 in Hela-si NT cells. Weakened band was observed when Hela-si Caspase-9 sample was tested. Hela-si NT and Hela-si Caspase-9 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1610-95, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

Fig5: Western blot analysis of Active+Pro Caspase-9 on different lysates with Rabbit anti-Active+Pro Caspase-9 antibody (R1308-12) at 1/1,000 dilution.

Lane 1: Hela-si NT cell lysate
Lane 2: Hela-si Caspase-9 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 46 kDa
Observed band size: 46 kDa

Exposure time: 5 minutes;

4-20% SDS-PAGE gel.

R1308-12 was shown to specifically react with Active+Pro Caspase-9 in Hela-si NT cells. Weakened band was observed when Hela-si Active+Pro Caspase-9 sample was tested. Hela-si NT and Hela-si Active+Pro Caspase-9 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (R1308-12, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.