Matrigengel Matrix GFR Phenol Red Free HC-082723

Description

PRODUCT DESCRIPTIONÂ

Basement membranes are continuous sheets of specialized extralar an interface between endothelial, epithelial, muscle, or neuronal and their adjacent stroma. Basement membranes are degraded and regenerated during development and wound healing. They not only support and layers, but they also play an essential role in tissue organization that affects adhesion, migration, proliferation, and differentiation. Basement membranes provide major barriers to invasion by metastatic tumor .

Wintop Matrigengel c14;ˆGFR nol Red Free HC）is a soluble form of basement membrane purified from Engelbreth-Holm-Swarm (EHS) tumor.Wintop Matrigengel c14;ˆGFR nol Red Free HC）at 37 °C to form a reconstituted basement membrane. The major components of Wintop Matrigengel c14;ˆGFR nol Red Free HC）include laminin, collagen IV, entactin, and heparin sulfate proteoglycan.

INTENDED USE

This series is divided into standard high concentration and low factor high concentration, their concentration and viscosity are higher than the ordinary concentration of ch is more suitable for in vivo experiments (animal models), such as PDX, CDX, in vitro angiogenesis assays.

MATERIALS PROVIDED

PRODUCT PARAMETER

Source: Mouse Tumor

Appearance：

â‘ Â Color:Â nol-containing red matrigengel is yellow-pink, and nol-free red anslucent light yellow;

② Form: standard matrigengel dissolved at 4℃, a transparent liquid state; High concentration matrigengel after 0℃ solution, transparent liquid state, 4℃ for a long time to show a semi-gel.

Concentration:Â Protein concentration ranges from 8 to 26mg/mL

Endo‰¤Â 4.5EU/ mL

Gel time: 5-30min gel at room temperature, the speed of gel formation is accelerated when the temperature is from 22°C to 37°C.

MATERIAL QUALIFICATIONS

l Routine screening of mouse colony pathogensby mouse antibody product (MAP) tests

l Testing for bacteria, fungi and mycoplasmato ensure negative results

l Extensive PCR testing for a variety of pathogens including LDEV to ensure strict control of raw materials used in the production process

l Extraction from LDEV-free mouse tumor

l Gel stability testing at 37℃ for 14 days

l Detection of endousing serological methods

l Biological function verification of each lot (organoid culture and differentiation experiments; Subcutaneous tumor formation test; Stem culture; Angiogenesis experiment etc.)

COATING PROCEDURES:

Thaw Matrigengel ernight at 2-8 °C. Refrigerator temperatures may vary, therefore it is recommended to keep Matrigengel on ice in a refrigerator during the thawing process. Thawed Matrigengel solidifies quickly at temperatures above 15 °C; when working with Matrigengel,

keep it on ice to prevent untimely gelling.

There are many applications for Matrigengel which require different thicknesses and concentrations. A thick gel is needed for applications such as endothelial formation of capillary-like structures (Tube Formation Assay), the differentiation of rat aorta tissue into capillary-like structures (Aortic Ring Assay), epithelial organoid formation, or tumor organoid formation. Some applications, such as propagation of primary , require a thin layer coating and not a thick gel; therefore, the thin layer method should be used.

Thick Gel Method:

Thin Layer Method (non-gelling):

Example for PROCEDURES:

Subcutaneous tumor formation in mice

1.1. Equipment: Biosafety cabinet, incubator, low temperature horizontal centrifuge, inverted microscope

1.2.Reagents:Matrigengel(Ca1),basic medium,medium containing 10% fetal bovine serum, 1×PBS, Trypsin Solution

1.3.Consumables: sterile pipette tips; 10cm culture dish; Sterile EP tube; Disposable syringes and other consumables can be adjusted according to the experimental design

2.1.Put the Matrigengel in the ice box and put it in the refrigerator at 4℃ so that the glue can slowly melt overnight;

2.2. Select in good condition and growth logarithmically, discard the supernatant, add 2 mL 1×PBS solution to wash gently, discard the liquid;

2.3. Add 1 mL 0.25% trypsin digestion solution into the petri dish, let it stand for 10 seconds, discard the trypsin, and continue digestion at room temperature for 1~3 minutes with the residual trypsin.

2.4. When the become round (keep in mind that they cannot be digested until the edge is clear), add 1 mL medium containing 10% fetal bovine serum to terminate digestion. Carefully blow the apart and collect the suspension into a 15 mL plastic centrifuge tube.

2.5. Centrifuge at 1000 rpm for 3 minutes and discard the supernatant. After washing the with PBS for one time, the were re-suspended by adding serum-free basal medium, and 10 μL suspension was taken for count.

2.6.

HepG2 : Each mice should be vaccinated 5 million . According to density, the required suspension was placed in a sterile 1.5mL plastic centrifuge tube at 1000 rpm for 3 min. The supernatant was discarded and the serum-free medium or PBS was added until the total volume was 300μL.

HCT – 116 :Each mice should be vaccinated 1 million . According to density, the required suspension was placed in a sterile 1.5mL plastic centrifuge tube at 1000 rpm for 3 min. The supernatant was discarded and the serum-free medium or PBS was added until the total volume was 300μL.

MIA – PaC- 2 : Each mice should be vaccinated 10 million . According to density, the required suspension was placed in a sterile 1.5mL plastic centrifuge tube at 1000 rpm for 3 min. The supernatant was discarded and the serum-free medium or PBS was added until the total volume was 500μL.

2.7.Mixing of the Matrigengel: suspension and Matrigengel are mixed in a 1:1 ratio at 4℃.

2.8.Subcutaneous injection: The nude mice were fixed with the left hand and injected subcutaneously into the right back of the nude mice. During inoculation, the needle was inserted a little deeper into the subcutaneously, about 1 cm deep, to reduce the overflow of suspension from the eye after injection, and the inoculation volume was 100μL. MIA-PaC-2 were injected 200 μL/ piece.

2.9. Recording data: The nude mice were put back into the cage for further feeding, the tumor volume was measured regularly according to the experimental requirements, the data was recorded to make a curve, and the nude mice were euthanized before the tumor volume was less than 2000 mm3, and the tumor was removed and tograd.