Mouse alpha smooth muscle Actin Ready-To-Use IHC Kit 

小鼠肌动蛋白α/α-SMA/α Actin单克隆即用型免疫组化试剂盒

Catalog Number: KL1IHC0114M

英文名称：Mouse alpha smooth muscle Actin Ready-To-Use IHC Kit

中文名称：小鼠肌动蛋白α/α-SMA/α Actin单克隆即用型免疫组化试剂盒

别名：alpha sarcomeric Actin; alpha smooth muscle Actin; Actin alpha; ASMA; ASM-A; alpha-SMA; alpha SMA; AAT6; ACTA2; Actin alpha 2 smooth muscle aorta; Actin aortic smooth muscle; ACTSA; ACTVS; Alpha 2 actin; Alpha-actin 2; Cell growth inhibiting gene 46 protein; Growth inhibiting gene 46; ACTA_HUMAN; Actin alpha 2 smooth muscle aorta; Actin aortic smooth muscle; Actin, aortic smooth muscle; Alpha 2 actin; Alpha actin 2; Alpha cardiac actin; Alpha-actin 2; Alpha-actin-2; Cell growth inhibiting gene 46 protein; Cell growth-inhibiting gene 46 protein; Growth inhibiting gene 46; MYMY5

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

Smooth Muscle Actin belongs to the actin family of proteins, which are highly conserved proteins that play a rolein cell motility, structure and integrity. Alpha, beta and gamma actin isoforms have been identified, with alphaactin being a major constituent of the contractile apparatus, while beta and gamma actins are involved in theregulation of cell motility. In particular, smooth muscle actin is an alpha actin that is found in skeletal muscle. Actin exists as a ubiquitous protein involved with filament formation that make up large portions of thecytoskeleton. Actin filaments interact with myosin to assist in muscle contraction as well as aiding in cell motilityand cytokinesis. Smooth muscle actin is found on smooth muscle vessel walls, gut wall, myometrium, myoepithelial cells in breast and salivary glands. Defects in the smooth muscle actin gene cause aorticaneurysm familial thoracic type 6. Actin isoforms differ slightly in their N-terminus and the sequences of each areperfectly conserved in higher vertebrates. Alpha-smooth muscle actin is abundant in vascular and visceralsmooth muscle cells. In addition, it has also been shown that smooth muscle actin appear in stress fibers offibroblastic cells during pathological situations involving contractile phenomena such as wound healing andfibrocontractive diseases. Multiple alternatively spliced variants of smooth muscle actin have been identified.

Synonyms

alpha sarcomeric Actin; alpha smooth muscle Actin; Actin alpha; ASMA; ASM-A; alpha-SMA; alpha SMA; AAT6;ACTA2; Actin alpha 2 smooth muscle aorta; Actin aortic smooth muscle; ACTSA; ACTVS; Alpha 2 actin;Alpha-actin 2; Cell growth inhibiting gene 46 protein; Growth inhibiting gene 46; ACTA_HUMAN; Actin alpha 2smooth muscle aorta; Actin aortic smooth muscle; Actin, aortic smooth muscle; Alpha 2 actin; Alpha actin 2;Alpha cardiac actin; Alpha-actin 2; Alpha-actin-2; Cell growth inhibiting gene 46 protein; Cell growth-inhibitinggene 46 protein; Growth inhibiting gene 46; MYMY5.

Validation Data

Immunohistochemical analysis of paraffin embedded mouse colon tissueslide 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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