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KD-Validated GOLPH3 Mouse mAb

#66159

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shRNA Knockdown Validated
Western blotting analysis using golgi phosphoprotein 3 antibody (Cat#66159). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with golgi phosphoprotein 3 antibody (Cat#66159, 1:2,500) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Western blotting analysis using golgi phosphoprotein 3 antibody (Cat#66159). Golgi phosphoprotein 3 expression in wild-type (WT) and golgi phosphoprotein 3 (GOLPH3) shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with golgi phosphoprotein 3 antibody (Cat#66159, 1:2,500) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
Immunocytochemical staining of HepG2 cells with Golgi phosphoprotein 3 antibody (Cat#66159, 1:1,000). Nuclei were stained blue with DAPI; Golgi phosphoprotein 3 was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Scale bar, 20 μm.
Flow cytometric analysis of Golgi phosphoprotein 3 expression in HepG2 cells using Golgi phosphoprotein 3 antibody (Cat#66159, 1:2,000). Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Green, isotype control; red, Golgi phosphoprotein 3.
Validation of Golgi phosphoprotein 3 knockdown using flow cytometry. Wild-type(WT, Blue) and Golgi phosphoprotein 3 knockdown(KD, Green) HeLa cells were stained with Golgi phosphoprotein 3 antibody(Cat#66159, 1:2,000) and analyzed using BD flow cytometer. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS.
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  • Gene Symbol:
    GOLPH3
  • Catalog:
    66159
  • Application:
    WB, FCM, ICC
  • Species Reactivity:
    H, M, R
Information
Product Name KD-Validated GOLPH3 Mouse mAb
Aliases GOLPH3; Golgi Phosphoprotein 3; GPP34; MIDAS; GOPP1; Vps74; Golgi Peripheral Membrane Protein 1, 34 KDa; Mitochondrial DNA Absence Factor; Golgi-Associated Protein; Coat Protein GPP34; Golgi Protein; Coat-Protein; Golgi Phosphoprotein 3 (Coat-Protein)
Background

Gene Name: GOLPH3

NCBI Gene Entry: 64083

UniProt Entry: Q9H4A6

Application Information

Molecular Weight: Predicted, 34 kDa; observed, 34 kDa

Clonality: Mouse monoclonal antibody

Clone ID: 25GB7340

Species Reactivity: Human, mouse, rat

Applications Tested: Western Blotting (WB), flow cytometry (FCM), immunocytochemistry (ICC)

Immunogen Purified protein of human GOLPH3
Isotype Mouse IgG1
Storage Buffer Supplied in PBS (pH 7.4) containing 0.02% sodium azide.
Storage Store at -20 °C for one year.
Recommended Dilutions Western Blotting (WB): 1:500-1:2,500
Flow Cytometry (FCM): 1:200-1:2,000
Immunocytochemistry (ICC): 1:100-1:1,000
Note This product is for research use only.
Data
  • Western blotting analysis using golgi phosphoprotein 3 antibody (Cat#66159). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with golgi phosphoprotein 3 antibody (Cat#66159, 1:2,500) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
  • Western blotting analysis using golgi phosphoprotein 3 antibody (Cat#66159). Golgi phosphoprotein 3 expression in wild-type (WT) and golgi phosphoprotein 3 (GOLPH3) shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with golgi phosphoprotein 3 antibody (Cat#66159, 1:2,500) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
  • Immunocytochemical staining of HepG2 cells with Golgi phosphoprotein 3 antibody (Cat#66159, 1:1,000). Nuclei were stained blue with DAPI; Golgi phosphoprotein 3 was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Scale bar, 20 μm.
  • Flow cytometric analysis of Golgi phosphoprotein 3 expression in HepG2 cells using Golgi phosphoprotein 3 antibody (Cat#66159, 1:2,000). Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Green, isotype control; red, Golgi phosphoprotein 3.
  • Validation of Golgi phosphoprotein 3 knockdown using flow cytometry. Wild-type(WT, Blue) and Golgi phosphoprotein 3 knockdown(KD, Green) HeLa cells were stained with Golgi phosphoprotein 3 antibody(Cat#66159, 1:2,000) and analyzed using BD flow cytometer. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS.
Western blotting analysis using golgi phosphoprotein 3 antibody (Cat#66159). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with golgi phosphoprotein 3 antibody (Cat#66159, 1:2,500) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Western blotting analysis using golgi phosphoprotein 3 antibody (Cat#66159). Golgi phosphoprotein 3 expression in wild-type (WT) and golgi phosphoprotein 3 (GOLPH3) shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with golgi phosphoprotein 3 antibody (Cat#66159, 1:2,500) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
Immunocytochemical staining of HepG2 cells with Golgi phosphoprotein 3 antibody (Cat#66159, 1:1,000). Nuclei were stained blue with DAPI; Golgi phosphoprotein 3 was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Scale bar, 20 μm.
Flow cytometric analysis of Golgi phosphoprotein 3 expression in HepG2 cells using Golgi phosphoprotein 3 antibody (Cat#66159, 1:2,000). Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Green, isotype control; red, Golgi phosphoprotein 3.
Validation of Golgi phosphoprotein 3 knockdown using flow cytometry. Wild-type(WT, Blue) and Golgi phosphoprotein 3 knockdown(KD, Green) HeLa cells were stained with Golgi phosphoprotein 3 antibody(Cat#66159, 1:2,000) and analyzed using BD flow cytometer. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS.
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