Description: Collagen type IV is a major component of basement membranes, such as those associated with capillaries and within glomeruli. This antibody will help detect collagen type IV and determine the presence of an intact basal lamina in normal tissues or the loss of structure as seen in various carcinomas.
Primary Antibody
Name: Collagen Type IV Antibody |
Clone: PHM-12, Mouse anti-Human |
Supplier: Vector Labs |
Catalog Number: VP-C388 |
Dilution: 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. |
Incubation Time/Temp: 60 minutes/room temperature |
Device: Steamer, Incubator |
Buffer/pH value: Citrate buffer/pH 6.0 for 20 minutes and let sections cool for 5-10 minutes. Then followed immediately by Proteinase K digestion for 30 seconds at 37 °C. Cool sections for another 10 minutes at room temperature. |
Heat/Cool Temperature: 95-100 ºC/room temperature for citrate buffer retrieval, 37ºC/room temperature for proteinase K digestion. |
Heat/Cool Time: follow instruction above |
Standard Method: ABC Method or LSAB Method |
Enhanced Method: Polymeric Methods |
Reagent: DAB |
Incubation Time/Temperature: 1-3 minutes/room temperature |
Reagent: Mayer's Hematoxylin |
Staining Time: 30 seconds |
Staining Pattern: Membrane |
Images: Search image |
Tissue Type: Kidney, skin |
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections |
Positive Control: Kidney, skin |
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 |
There is no need to perform antigen retrieval on frozen sections |
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