ER Stress-induced Autophagy Antibody Sampler Kit
cat.: HAK21029
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: P11021 Human | P20029 Mouse | P06761 Rat | P05198 Human | Q6ZWX6 Mouse | P68101 Rat | P05198 Human | Q6ZWX6 Mouse | P68101 Rat | O94817 Human | Q14457 Human | O88597 Mouse | Q91XJ1 Rat | P45983 Human | P45984 Human | P53779 Human | Q61831 Mouse | Q91Y86 Mouse | Q9WTU6 Mouse | P49185 Rat | P49186 Rat | P49187 Rat | P45983 Human | P45984 Human | P53779 Human | Q61831 Mouse | Q91Y86 Mouse | Q9WTU6 Mouse | P49185 Rat | P49186 Rat | P49187 Rat
Images
HAK21029_1.jpg Fig1: Western blot analysis of GRP78 BiP on different lysates with Mouse anti-GRP78 BiP antibody (HA601076) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate (10 µg/Lane)
Lane 2: MCF-7 cell lysate (10 µg/Lane)
Lane 3: RAW264.7 cell lysate (10 µg/Lane)
Lane 4: F9 cell lysate (10 µg/Lane)
Lane 5: NIH/3T3 cell lysate (10 µg/Lane)
Lane 6: PC-12 cell lysate (10 µg/Lane)
Lane 7: PMVEC cell lysate (10 µg/Lane)
Lane 8: Mouse brain tissue lysate (20 µg/Lane)

Predicted band size: 72 kDa
Observed band size: 75 kDa

Exposure time: 1 minute;

8% 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 (HA601076) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.
HAK21029_2.jpg Fig2: Western blot analysis of EIF2S1 on different lysates with Rabbit anti-EIF2S1 antibody (HA500385) at 1/500 dilution.

Lane 1: Mouse colon tissue lysate, 20 µg/Lane
Lane 2: Rat spleen tissue lysate, 20 µg/Lane

Predicted band size: 36 kDa
Observed band size: 36 kDa

Exposure time: 30 seconds;

12% 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 (HA500385) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
HAK21029_3.jpg Fig3: Western blot analysis of Phospho-EIF2S1 (S51) on different lysates with Rabbit anti-Phospho-EIF2S1 (S51) antibody (ET1603-14) at 1/2,000 dilution.

Lane 1: HeLa whole cell lysate (15 µg/Lane)
Lane 2: HeLa treated with 50nM Calyculin A for 3 hours whole cell lysate (15 µg/Lane)
Lane 3: NIH/3T3 whole cell lysate (15 µg/Lane)
Lane 4: NIH/3T3 treated with 100nM Calyculin A for 30 minutes whole cell lysate (15 µg/Lane)
Lane 5: C6 whole cell lysate (20 µg/Lane)
Lane 6: C6 treated with 100nM Calyculin A for 30 minutes whole cell lysate (20 µg/Lane)

Predicted band size: 36 kDa
Observed band size: 36 kDa

Exposure time: Lane 1-4: 2 minutes; Lane 5-6: 23 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 (ET1603-14) 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.
HAK21029_4.jpg Fig4: Western blot analysis of ATG12 on different lysates with Rabbit anti-ATG12 antibody (HA721504) at 1/1,000 dilution.

Lane 1: HeLa cell lysate, 20 µg/Lane
Lane 2: HCT 116 cell lysate, 20 µg/Lane
Lane 3: HEK-293 cell lysate, 20 µg/Lane
Lane 4: SH-SY5Y cell lysate, 20 µg/Lane

Predicted band size: 15 kDa
Observed band size: 55/20 kDa

Exposure time: 3 minutes;

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 (HA721504) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
HAK21029_5.jpg Fig5: Western blot analysis of Beclin 1 on different lysates with Rabbit anti-Beclin 1 antibody (HA721216) at 1/500 dilution.

Lane 1: Rat cerebellum tissue lysate, 20 µg/Lane
Lane 2: Mouse cerebellum tissue lysate, 20 µg/Lane

Predicted band size: 52 kDa
Observed band size: 52 kDa

Exposure time: 2 minutes;

10% 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 (HA721216) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
HAK21029_6.jpg Fig6: Western blot analysis of JNK1+JNK2+JNK3 on different lysates with Rabbit anti-JNK1+JNK2+JNK3 antibody (ET1601-28) at 1/2,000 dilution and competitor's antibody at 1/1,000 dilution.

Lane 1: HeLa cell lysate, 15 µg/Lane
Lane 2: HEK-293 cell lysate, 15 µg/Lane
Lane 3: Jurkat cell lysate, 15 µg/Lane
Lane 4: MCF7 cell lysate, 15 µg/Lane
Lane 5: NIH/3T3 cell lysate, 15 µg/Lane
Lane 6: PC-12 cell lysate, 15 µg/Lane
Lane 7: C6 cell lysate, 15 µg/Lane

Predicted band size: 48/53 kDa
Observed band size: 48/53 kDa

Exposure time: 35 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 (ET1601-28) 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.
HAK21029_7.jpg Fig7: Western blot analysis of Phospho-JNK1/2/3(T183+T183+T221) on different lysates with Rabbit anti-Phospho-JNK1/2/3(T183+T183+T221) antibody (ET1609-42) at 1/1,000 dilution.

Lane 1: Hela cell lysate, untreated, 10 µg/Lane
Lane 2: Hela cell lysate, treated with Anisomycin, 10 µg/Lane
Lane 3: A431 cell lysate, untreated, 10 µg/Lane
Lane 4: A431 cell lysate, treated with UV40, 10 µg/Lane
Lane 5: 293 cell lysate, untreated, 10 µg/Lane
Lane 6: 293 cell lysate, treated with UV40, 10 µg/Lane
Lane 7: Hela cell lysate, untreated, 10 µg/Lane
Lane 8: Hela cell lysate, treated with UV40, 10 µg/Lane

Predicted band size: 48/53 kDa
Observed band size: 48/53 kDa

Exposure time: 2 minutes;

12% 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 (ET1609-42) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 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”.