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 | P97864 Mouse
Entrez Gene: 64026 Rat
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
K2001_1.jpg 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.
K2001_2.jpg 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.
K2001_3.jpg 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.
K2001_4.jpg 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.
K2001_5.jpg 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.
K2001_6.jpg Fig6: Western blot analysis of Caspase-7 on different lysates with Rabbit anti-Caspase-7 antibody (HA723558) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 1μM staurosporine for 3 hours cell lysate
Lane 3: C2C12 cell lysate
Lane 4: C2C12 treated with 1μM staurosporine for 3 hours cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 34 kDa
Observed band size: 34/32/20/18 kDa

Exposure time: 2 minute 30 seconds; ECL: K1801;

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 (HA723558) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
K2001_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-Caspase-7 antibody (HA723558) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723558) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.