LS-C756473

antibody from LSBio

Targeting: GOLPH3 GOPP1, GPP34, MIDAS, Vps74

Western blot (Enhanced validation)

ELISA (Recommended by provider)

Immunohistochemistry (Enhanced validation)

Antibody Data

Product number

LS-C756473

Provider

LSBio

Product name

GOLPH3 Antibody LS-C756473

Antibody type

Polyclonal

Description

Immunogen affinity purified

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Storage

After reconstitution, may be stored at 4°C for 1 month. For long-term storage and to avoid freeze-thaw cycles, aliquot and store at -20°C.

Western blot

Enhanced validation

Submitted by

LSBio (provider)

Enhanced method

Genetic validation

Main image

Experimental details

Western blot analysis of GOLPH3 using anti-GOLPH3 antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: human Hela whole cell lysate,Lane 2: human placenta tissue lysates,Lane 3: human U20S whole cell lysate,Lane 4: human Caco-2 whole cell lysate,Lane 5: human SW620 whole cell lysate,Lane 6: human A549 whole cell lysate. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GOLPH3 antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for GOLPH3 at approximately 34KD. The expected band size for GOLPH3 is at 34KD.

Enhanced validation

Submitted by

LSBio (provider)

Enhanced method

Genetic validation

Main image

Experimental details

Western blot analysis of GOLPH3 using anti-GOLPH3 antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions. Lane 1: rat heart tissue lysates,Lane 2: rat brain tissue lysates,Lane 3: rat stomach tissue lysates,Lane 4: rat testis tissue lysates,Lane 5: mouse heart tissue lysates,Lane 6: mouse brain tissue lysates,Lane 7: mouse stomach tissue lysates,Lane 8: mouse testis tissue lysates,Lane 9: mouse NIH3T3 whole cell lysate. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GOLPH3 antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for GOLPH3 at approximately 34KD. The expected band size for GOLPH3 is at 34KD.

Immunohistochemistry

Enhanced validation

Submitted by

LSBio (provider)

Enhanced method

Genetic validation

Main image

Experimental details

IHC analysis of GOLPH3 using anti-GOLPH3 antibody. GOLPH3 was detected in frozen section of human placenta tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1µg/ml rabbit anti-GOLPH3 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

LSBio (provider)

Enhanced method

Genetic validation

Main image

Experimental details

IHC analysis of GOLPH3 using anti-GOLPH3 antibody. GOLPH3 was detected in frozen section of human placenta tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1µg/ml rabbit anti-GOLPH3 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

LSBio (provider)

Enhanced method

Genetic validation

Main image

Experimental details

IHC analysis of GOLPH3 using anti-GOLPH3 antibody. GOLPH3 was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2µg/ml rabbit anti-GOLPH3 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

LSBio (provider)

Enhanced method

Genetic validation

Main image

Experimental details

IHC analysis of GOLPH3 using anti-GOLPH3 antibody. GOLPH3 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2µg/ml rabbit anti-GOLPH3 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

LSBio (provider)

Enhanced method

Genetic validation

Main image

Experimental details

IHC analysis of GOLPH3 using anti-GOLPH3 antibody. GOLPH3 was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2µg/ml rabbit anti-GOLPH3 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

LSBio (provider)

Enhanced method

Genetic validation

Main image

Experimental details

IHC analysis of GOLPH3 using anti-GOLPH3 antibody. GOLPH3 was detected in paraffin-embedded section of rat small intestine tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2µg/ml rabbit anti-GOLPH3 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

LSBio (provider)

Enhanced method

Genetic validation

Main image

Experimental details

IHC analysis of GOLPH3 using anti-GOLPH3 antibody. GOLPH3 was detected in paraffin-embedded section of mouse lung tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1µg/ml rabbit anti-GOLPH3 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

LSBio (provider)

Main image

Experimental details

IHC analysis of GOLPH3 using anti-GOLPH3 antibody. GOLPH3 was detected in paraffin-embedded section of mouse lung tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1µg/ml rabbit anti-GOLPH3 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

LSBio (provider)

Main image

Experimental details

IHC analysis of GOLPH3 using anti-GOLPH3 antibody. GOLPH3 was detected in frozen section of human placenta tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1µg/ml rabbit anti-GOLPH3 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

LSBio (provider)

Main image

Experimental details

IHC analysis of GOLPH3 using anti-GOLPH3 antibody. GOLPH3 was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2µg/ml rabbit anti-GOLPH3 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

LSBio (provider)

Main image

Experimental details

IHC analysis of GOLPH3 using anti-GOLPH3 antibody. GOLPH3 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2µg/ml rabbit anti-GOLPH3 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

LSBio (provider)

Main image

Experimental details

IHC analysis of GOLPH3 using anti-GOLPH3 antibody. GOLPH3 was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2µg/ml rabbit anti-GOLPH3 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Supportive validation

Submitted by

LSBio (provider)

Main image

Experimental details

IHC analysis of GOLPH3 using anti-GOLPH3 antibody. GOLPH3 was detected in paraffin-embedded section of rat small intestine tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2µg/ml rabbit anti-GOLPH3 Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.