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SDF-1 Antibody Staining Protocol for Immunohistochemistry
Description: The C-X-C or α chemokine family is characterized by a pair of cysteine residues separated by a single amino acid and primarily functions as chemoattractants for neutrophils. The C-X-C family includes IL-8, NAP-2, MSGA and stromal cell-derived factor-1, or SDF-1. SDF-1 was originally described as a pre-B cell stimulatory factor, but has now been shown to function as a potent chemoattractant for T cells and monocytes, but not neutrophils. Receptors for the C-X-C family are G protein-coupled, seven- pass, transmembrane domain proteins which include IL-8RA, IL-8RB and fusin (also designated LESTR or CXCR-4). Fusin is highly homologous to the IL-8 receptors, sharing 37% sequence identity at the amino acid level. The IL-8 receptors bind to IL-8, NAP-2 and MSGA, while fusin binds to its cognate ligand, SDF-1. Fusin has been identified as the major co-receptor for T-tropic HIV-1, and SDF-1 has been shown to inhibit HIV-1 infection. Six human SDF-1 isoforms exist due to alternative splicing of CXCL12, the gene encoding SDF-1. Three isoforms are known for mouse and rat.
Primary Antibody
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Name: SDF-1 (C-19) Antibody |
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Clone: Goat polyclonal |
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Supplier: Santa Cruz Biotechnology |
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Catalog Number: sc-6193 |
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Dilution: 1:50 - 1:100 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: Cytoplasmic |
| Images: Search image |
Additional Information:
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Species Reactivity: Mouse, rat, human |
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Fixation: Formalin fixed, paraffin embedded sections |
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Positive Control: Skin |
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Negative Control: Omit primary antibody, isotype control or absorption control |
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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 |
References: