ICAM3 Recombinant Rabbit Monoclonal Antibody [PSH20-97]
cat.: HA724192
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, IHC-P, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH20-97 |
| 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: 60 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human ICAM3 aa 1-485. |
| Positive control: | U-937 (Human acute monocytic leukemia cells) cell lysate, Jurkat (Human T-lymphoblastic cells) cell lysate, Ramos (Human Burkitt's lymphoma cells) cell lysate, THP-1 (Human acute monoblastic leukemia cells) cell lysate, HL-60 (Human acute promyelocytic leukemia cells) cell lysate. |
| Subcellular location: | Membrane. |
| Recommended Dilutions:
WB IF-Cell IHC-P FC IP |
1:5,000 1:500 1:1,000 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P32942 Human |
| Alternative names: | CD50 CDW 50 CDw50 ICAM 3 ICAM-3 ICAM-R ICAM3 ICAM3_HUMAN ICAMR Intercellular adhesion molecule 3 |
Images
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Fig1:
Western blot analysis of ICAM3 on different lysates with Rabbit anti-ICAM3 antibody (HA724192) at 1/5,000 dilution. Lane 1: U-937 (Human acute monocytic leukemia cells) cell lysate Lane 2: Jurkat (Human T-lymphoblastic cells) cell lysate Lane 3: Ramos (Human Burkitt's lymphoma cells) cell lysate Lane 4: THP-1 (Human acute monoblastic leukemia cells) cell lysate Lane 5: HL-60 (Human acute promyelocytic leukemia cells) cell lysate Lysates/proteins at 15 µg/Lane. Exposure time: 20 seconds; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA724192, 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: 60 kDa Observed band size: 100-150 kDa |
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Fig2:
Western blot analysis of ICAM3 on different lysates with Rabbit anti-ICAM3 antibody (HA724192) at 1/5,000 dilution. Lane 1: Jurkat (Human T-lymphoblastic cells) cell lysate Lane 2: Jurkat treated with PNGseF cell lysate Lane 3: U-937 (Human acute monocytic leukemia cells) cell lysate Lane 4: U-937 treated with PNGseF cell lysate Lysates/proteins at 10 µg/Lane. Exposure time: 3 minutes; ECL: K1801 Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: HA724192, 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: 60 kDa Observed band size: 100-150/60 kDa |
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Fig3:
Application: Immunocytochemistry (IF-cell) Species: Human Sample: Jurkat (Human T-lymphoblastic cells) 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: HA724192, 1/500, overnight at 4℃. 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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Fig4:
Application: Immunocytochemistry (IF-cell) Species: Human Sample: U-937 (Human acute monocytic leukemia cells) 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: HA724192, 1/500, overnight at 4℃. 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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Fig5:
Application: Immunohistochemistry (IHC-P) Species: Human Tissue: Tonsil Sample: Paraffin-embedded section Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃. 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: HA724192, 1/1,000, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
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Fig6:
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℃. 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: HA724192, 1/1,000, 1 hour at room temperature. Secondary antibody: HA1119, 20 minutes at room temperature. |
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Fig7:
Application: Flow Cytometry Species: Human Sample: Jurkat (Human T-lymphoblastic cells) Blocking: 1% BSA + 10% normal goat serum + human FcR (Miltenyi Biotec), 15 minutes at room temperature. (human) Antibody dilution buffer: 1x PBS. Primary antibody: HA724192 (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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Fig8:
Application: Flow Cytometry Species: Human Sample: U-937 (Human acute monocytic leukemia cells) Blocking: 1% BSA + 10% normal goat serum + human FcR (Miltenyi Biotec), 15 minutes at room temperature. (human) Antibody dilution buffer: 1x PBS. Primary antibody: HA724192 (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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Fig9:
Immunoprecipitation (IP) ICAM3 was immunoprecipitated in 0.2 mg U-937 (Human acute monocytic leukemia cells) cell lysate with HA724192 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724192 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: U-937 cell lysate (input) Lane 2: HA724192 IP in U-937 cell lysate Lane 3: Rabbit IgG instead of HA724192 in U-937 cell lysate Exposure time: 14 seconds Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary dilution: HA724192, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature Predicted band size: 60 kDa Observed band size: 100-150 kDa |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.