Human Vimentin Ready-To-Use IHC Kit 

人波形蛋白单克隆即用型免疫组化试剂盒

Catalog Number: KL1IHC0113H

英文名称：Human Vimentin Ready-To-Use IHC Kit

中文名称：人波形蛋白单克隆即用型免疫组化试剂盒

别名：FLJ36605; OTTHUMP00000019224; VIM; VIME_HUMAN; Vimentin.

Sample Type: FFPE tissue

Size: 50T (including three control slides)

Storage and Stability: Please store components at the temperatures indicated on the individual tubelabels. The kit is stable for 6 months from the date of receipt.
General Information

Background

Vimentin is a developmentally regulated intermediate filament protein (IFP) found in cells of mesenchymal origin.It is believed to be involved with the intracellular transport of proteins between the nucleus and plasmamembrane. Unlike other IFP proteins, vimentin is expressed, along with desmin, during the early stages ofcellular development. During the development process, vimentin is exchanged for new, tissue-specific IFPs. Vimentin has been implicated to be involved in the rate of steroid synthesis via its role as a storage network forsteroidogenic cholesterol containing lipid droplets. Vimentin phosphorylation by a protein kinase causes thebreakdown of intermediate filaments and activation of an ATP and myosin light chain dependent contractile event. This results in cytoskeletal changes that facilitate the interaction of the lipid droplets within mitochondria, andsubsequent transport of cholesterol to the organelles leading to an increase in steroid synthesis.

Synonyms

FLJ36605; OTTHUMP00000019224; VIM; VIME_HUMAN; Vimentin

Validation Data

Immunohistochemical analysis of paraffin embedded human cervicalcarcinoma tissue slide using

Immunohistochemistry Protocol

1. Deparaffinization And Rehydration
Immerse slides in fresh xylene for 15 minutes and then repeat two more times using separate containers.Immerse slides sequentially in 100%, 95%, 90%, 80%, and 70% ethanol solutions for 5 minutes each. Rinseslides 3 times with distilled water for 5 minutes each.
2. Antigen Retrieval
Add 100×Antigen Retrieval Buffer into distilled water to prepare a 1×solution. Boil slides in 1×solution at95°C-100°C for 15 minutes. Move the slides to 1×solution at room temperature (RT) and allow them to stand for20 minutes. Rinse 3 times with PBS Buffer (dissolve the powder in 2L distilled water) for 5 minutes each.
3. Block Endogenous Peroxidase
Drain the liquid off the slides and then use a hydrophobic IHC pen to draw circles on the slides around tissuesections. Add 2-4 drops of Endogenous Peroxidase Blocking Buffer directly on slides, covering the wholetissue and block slides for 15 minutes at RT. Rinse 3 times with PBS Buffer for 5 minutes each.
4. Serum Blocking
Block with 2-4 drops of Blocking Buffer for 20 minutes at RT.
5. Primary Antibody Incubation
Drain blocking buffer from slides. Incubate slides with 2-4 drops of Histone H3 Mouse mAb overnight at 4°C or1-2 hours at RT. Rinse 3 times with PBS Buffer for 5 minutes each.

6. Secondary Antibody Incubation
Incubate slides with 2-4 drops of HRP-Goat anti-Mouse IgG pAb for 1-2 hours at RT. Rinse slides 3 times withPBS Buffer for 5 minutes each.
7. Signal Development
Remove residual liquid around the tissue section. Add 50ul fresh DAB Buffer (Chromogen Component A :Chromogen Component B : PBS Buffer=1:1:18) to cover the tissue. Monitor the reaction under themicroscope until a brown color is visible (approximate 3-5 minutes at RT). Stop reaction immediately by rinsingwith distilled water. Rinse slides 3 times with distilled water for 5 minutes each.
8. Counterstain
Counterstain with an appropriate amount of Counter Staining Reagent for 3-5 minutes at RT. Rinse slides withdistilled water for 5 minutes. Use 2-4 drops of Differentiation reagent to cover the tissue for 30 seconds. Rinseslides twice with distilled water for 5 minutes each.
9. Dehydration Sheet
Immerseslides sequentially in 70%, 80%, 90%, 95%, and 100% ethanol for 5 minutes each at RT. Immerseslides in 2 changes of fresh xylene, 15 minutes each. Drop some Mounting Media on the tissue. Mountcoverslips.
Notes
1. The positive control slide provided in the kit allows you to be sure that the experimental set-up is workingproperly.
2. Do not allow slides to dry at any time during this procedure.
3. Please don't replace the matching reagents in this product with other manufacturers' products.
4. As DAB is a carcinogen, please take necessary precautions.
5. PBS (reagent 1) can be stored for one week at 4℃ after preparation; The antigen retrieval buffer (1×reagent2) and the chromogenic agent (the mixture of reagents 7 and 8) should be prepared right before each assay.
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