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Western blot analysis of DNAJC5 on different lysates with Rabbit anti-DNAJC5 antibody (HA724445) at 1/5,000 dilution.
Lane 1: HeLa (Human cervical adenocarcinoma cells) cell lysate
Lane 2: THP-1 (Human acute monoblastic leukemia cells) cell lysate
Lane 3: HepG2 (Human liver cancer cells) cell lysate
Lane 4: Huh7 (Human liver cancer cells) cell lysate
Lane 5: NIH/3T3 (Mouse fibroblasts) cell lysate
Lane 6: RAW264.7 (Mouse monocytic macrophage leukemia cells) cell lysate
Lane 7: PC-12 (Rat pheochromocytoma cells (undifferentiated)) cell lysate
Lane 8: C6 (Rat glioma cells) cell lysate
Lane 9: COS-1 (African green monkey kidney fibroblasts) cell lysate
Lysates/proteins at 15 µg/Lane.
Exposure time: 7 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724445, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 22 kDa
Observed band size: 34 kDa
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Application: Immunocytochemistry (IF-cell)
Species: Rat
Sample: PC-12 (Rat pheochromocytoma cells (undifferentiated))
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: HA724445, 1/100, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).
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☑ Relative expression (RE)
Western blot analysis of DNAJC5 on different lysates with Rabbit anti-DNAJC5 antibody (HA724445) at 1/5,000 dilution.
Lane 1: Mouse brain tissue lysate
Lane 2: Mouse kidney tissue lysate
Lane 3: Rat brain tissue lysate
Lane 4: Rat kidney tissue lysate
Lane 5: Pig brain tissue lysate
Lysates/proteins at 30 µg/Lane.
Exposure time: 7 seconds; ECL: K1801
Negative expression of DNAJC5 protein in kidney tissue is consistent with the predicted expression pattern.
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724445, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 22 kDa
Observed band size: 34 kDa
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Application: Immunofluorescence (IHC-Fr)
Species: Rat
Tissue: Brain
Sample: Frozen section
Antigen retrieval: Not required
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: HA724445, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
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Application: Immunofluorescence (IHC-Fr)
Species: Mouse
Tissue: Brain
Sample: Frozen section
Antigen retrieval: Not required
Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: HA724445, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
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Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Placenta
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: HA724445, 1/200, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Brain
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: HA724445, 1/200, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Immunohistochemistry (IHC-P)
Species: Human
Tissue: Lung
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: HA724445, 1/200, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Immunohistochemistry (IHC-P)
Species: Mouse
Tissue: Placenta
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: HA724445, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Immunohistochemistry (IHC-P)
Species: Mouse
Tissue: Brain
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: HA724445, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Immunohistochemistry (IHC-P)
Species: Mouse
Tissue: Lung
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: HA724445, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Immunohistochemistry (IHC-P)
Species: Rat
Tissue: Placenta
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: HA724445, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Immunohistochemistry (IHC-P)
Species: Rat
Tissue: Brain
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: HA724445, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Immunohistochemistry (IHC-P)
Species: Rat
Tissue: Lung
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: HA724445, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
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Application: Flow Cytometry (Intra)
Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA724445 (1/1,000) (Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
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Application: Flow Cytometry (Intra)
Species: Mouse
Sample: NIH/3T3 (Mouse fibroblasts)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA724445 (1/1,000) (Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
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Application: Flow Cytometry (Intra)
Species: Rat
Sample: PC-12 (Rat pheochromocytoma cells (undifferentiated))
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: HA724445 (1/1,000) (Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
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Immunoprecipitation (IP)
DNAJC5 was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate with HA724445 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724445 at 1/25,000 dilution. Alpaca anti-Rabbit IgG Fc, Recombinant VHH monoclonal Antibody(HA1031) at 1/50000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input)
Lane 2: HA724445 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA724445 in HeLa cell lysate
Exposure time: 2 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary dilution: HA724445, 1/25,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Predicted band size: 22 kDa
Observed band size: 34 kDa
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