Endosomal Marker Antibody Sampler Kit

Western blot analysis of Clathrin heavy chain on different lysates with Rabbit anti-Clathrin heavy chain antibody (ET1704-50) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: A431 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: PC-12 cell lysate
Lane 5: HUVEC cell lysate
Lane 6: SH-SY5Y cell lysate
Lane 7: COS-1 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 180kDa
Observed band size: 180 kDa

Exposure time: 10 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 (ET1704-50) at 1/1,000 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.

Western blot analysis of Caveolin-1 on different lysates with Rabbit anti-Caveolin-1 antibody (ET1603-1) at different dilutions.

Lane 1/2: A549 cell lysate
Lane 3/4: A431 cell lysate

Lysates/proteins at 10 µg/Lane.
Predicted band size: 20 kDa
Observed band size: 20 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 (ET1603-1) at different dilutions were 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.

All lanes: Western blot analysis of RAB7 with anti-RAB7 antibody (ET1611-96) at 1:500 dilution.
Lane 1: Wild-type Hela whole cell lysate (10 µg).
Lane 2/3: RAB7 knockdown Hela whole cell lysate (10 µg).

ET1611-96 was shown to specifically react with RAB7 in wild-type Hela cells. Weakened bands were observed when RAB7 knockdown samples were tested. Wild-type and RAB7 knockdown 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 (ET1611-96, 1:500) was used in 5% BSA 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.

Western blot analysis of RAB7 on different lysates with Rabbit anti-RAB7 antibody (ET1611-96) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: MCF7 cell lysate
Lane 3: Neuro-2a cell lysate
Lane 4: C2C12 cell lysate
Lane 5: PC-12 cell lysate
Lane 6: C6 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 24 kDa
Observed band size: 24 kDa

Exposure time: 17 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 (ET1611-96) at 1/5,000 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.

Western blot analysis of Rab11A on different lysates with Rabbit anti-Rab11A antibody (HA721552) at 1/1,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: A549 cell lysate (20 µg/Lane)
Lane 3: SH-SY5Y cell lysate (20 µg/Lane)
Lane 4: Neuro-2a cell lysate (20 µg/Lane)
Lane 5: C6 cell lysate (20 µg/Lane)
Lane 6: Mouse testis tissue lysate (40 µg/Lane)
Lane 7: Rat testis tissue lysate (40 µg/Lane)
Lane 8: Rat brain tissue lysate (40 µg/Lane)

Predicted band size: 24 kDa
Observed band size: 24 kDa

Exposure time: 2 minutes 37 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 (HA721552) 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 1:100,000 dilution was used for 1 hour at room temperature.