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Flk-1 Antibody Staining Protocol for Immunohistochemistry
Description: Three cell membrane receptor tyrosine kinases, Flt (also designated VEGF-R1), Flk-1 (also designated VEGF-R2) and Flt-4, putatively involved in the growth of endothelial cells, are characterized by the presence of seven immunoglobulin-like sequences in their extracellular domain. These receptors exhibit high degrees of sequence relatedness to each other as well as lesser degrees of relatedness to the class III receptors including CSF-1/Fms, PDGR, SLFR/Kit and Flt-3/Flk-2. Two members of this receptor class, Flt-1 and Flk-1, have been shown to represent high affinity receptors for vascular endothelial growth factors (VEGFs). On the basis of structural similarity to Flt and Flk-1, it has been speculated that Flt-4 might represent a third receptor for either VEGF or a VEGF-related ligand.
Primary Antibody
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Name: Flk-1 Antibody |
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Clone: A-3, Mouse anti-Human |
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Supplier: Santa Cruz Biotechnology |
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Catalog Number: sc-6251 |
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Dilution: 1:50 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed. |
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Incubation Time/Temp: 60 minutes/room temperature |
Antigen Retrieval
| Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102) |
| Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100) |
| Heat/Cool Temperature: 95 ºC - 100 ºC/room temperature |
| Heat/Cool Time: 20 minutes/20 minutes |
Detection Methods
| Standard Method: ABC Method or LSAB Method |
| Enhanced Method: Polymeric Methods |
Chromogen Substrate
| Reagent: DAB |
| Incubation Time/Temperature: 1-3 minutes/room temperature |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Membrane |
| Images: Search image |
Additional Information:
| Tissue Type: Human colon carcinoma |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Human colon carcinoma |
| Negative Control: Omit primary antibody, isotype control or absorption control |
| Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary |
Notes:
Not tested on frozen sections |
References: