Phospho-SAPK/JNK (T183/Y185) Recombinant Rabbit Monoclonal Antibody [PSH24-76]
cat.: HA724473
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH24-76 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage buffer: | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 48 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic phospho-peptide corresponding to residues surrounding Thr183/Tyr185 of Human SAPK/JNK aa 161-204. |
| Positive control: | HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate, NIH/3T3 treated with 25μg/mL anisomycin for 30 minutes cell lysate, C6 treated with 25μg/mL anisomycin for 30 minutes cell lysate. |
| Subcellular location: | Cytoplasm, Nucleus, Synapse. |
| Recommended Dilutions:
WB IHC-P IP |
1:5,000 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P45983 Human |
Images
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Fig1:
Western blot analysis of Phospho-SAPK/JNK (T183/Y185) on different lysates with Rabbit anti-Phospho-SAPK/JNK (T183/Y185) antibody (HA724473) at 1/5,000 dilution.
Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate Lane 2: HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate Lane 3: HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour Lysates/proteins at 15 µg/Lane. Exposure time: 1minute 50 seconds; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA724473, 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: 48 kDa Observed band size: 42-54 kDa |
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Fig2:
Western blot analysis of Phospho-SAPK/JNK (T183/Y185) on different lysates with Rabbit anti-Phospho-SAPK/JNK (T183/Y185) antibody (HA724473) at 1/5,000 dilution.
Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate Lane 2: HeLa treated with 25μg/mL anisomycin for 30 minutes cell lysate Lane 3: NIH/3T3 (Mouse fibroblast) cell lysate Lane 4: NIH/3T3 treated with 25μg/mL anisomycin for 30 minutes cell lysate Lane 5: C6 (Rat glioma cell) cell lysate Lane 6: C6 treated with 25μg/mL anisomycin for 30 minutes cell lysate Lysates/proteins at 15 µg/Lane. Exposure time: 3 minutes; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA724473, 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: 48 kDa Observed band size: 42-54 kDa |
|
Fig3:
Application: Immunohistochemistry (IHC-P)
Species: Human Tissue: Breast carcinoma 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: HA724473, 1/1,000, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
|
Fig4:
Application: Immunohistochemistry (IHC-P)
Species: Human Tissue: Lung carcinoma 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: HA724473, 1/1,000, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
|
Fig5:
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: HA724473, 1/1,000, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
|
Fig6:
Application: Immunohistochemistry (IHC-P)
Species: Human Tissue: Colon (Myenteric plexus) 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: HA724473, 1/1,000, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
|
Fig7:
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: HA724473, 1/1,000, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
|
Fig8:
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: HA724473, 1/1,000, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
|
Fig9:
Immunoprecipitation (IP)
Phospho-SAPK/JNK (T183/Y185) was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) treated with 25µg/mL anisomycin for 30 minutes cell lysate with HA724473 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724473 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 treated with 25µg/mL anisomycin for 30 minutes cell lysate (input) Lane 2: HA724473 IP in HeLa treated with 25µg/mL anisomycin for 30 minutes cell lysate Lane 3: Rabbit IgG instead of HA724473 in HeLa treated with 25µg/mL anisomycin for 30 minutes cell lysate Exposure time: 46 seconds Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary dilution: HA724473, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature Predicted band size: 48 kDa Observed band size: 42-54 kDa |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.