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Western blot analysis of GPI on different lysates with Rabbit anti-GPI antibody (HA724519) at 1/5,000 dilution.
Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
Lane 2: HCT 116 (Human colon cancer cell) cell lysate
Lane 3: PC-3 (Human prostate cancer cell) cell lysate
Lane 4: HEK-293 (Human embryonic kidney cell) cell lysate
Lane 5: A549 (Human lung adenocarcinoma cell) cell lysate
Lane 6: COS-1 (African green monkey kidney fibroblast) cell lysate
Lysates/proteins at 10 µg/Lane.
Exposure time: 10 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724519, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 63 kDa
Observed band size: 55 kDa
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Western blot analysis of GPI on different lysates with Rabbit anti-GPI antibody (HA724519) at 1/5,000 dilution.
Lane 1: Mouse liver tissue lysate
Lane 2: Mouse spleen tissue lysate
Lane 3: Mouse brain tissue lysate
Lane 4: Rat liver tissue lysate
Lane 5: Rat spleen tissue lysate
Lane 6: Rat brain tissue lysate
Lysates/proteins at 20 µg/Lane.
Exposure time: 10 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724519, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 63 kDa
Observed band size: 55 kDa
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Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Colon
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724519, 1/500, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Kindey
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724519, 1/2,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Immunohistochemistry (IHC-P)
Species: Mouse
Tissue: Colon
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724519, 1/500, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Immunohistochemistry (IHC-P)
Species: Mouse
Tissue: Kindey
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724519, 1/2,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Immunohistochemistry (IHC-P)
Species: Rat
Tissue: Colon
Sample: Paraffin-embedded section
Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C.
Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA724519, 1/500, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Immunoprecipitation (IP)
GPI was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate with HA724519 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724519 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input)
Lane 2: HA724519 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA724519 in HeLa cell lysate
Exposure time: 3 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary dilution: HA724519, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Predicted band size: 63 kDa
Observed band size: 55 kDa
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