We use matrigel diluted in DMEM. We add 10ml of cold DMEM to the 10ml matrigel vial for a 1:1 dilution. These can then be aliquoted and frozen. We then dilute the aliquots 1:15, again in cold DMEM, so that the final working ration is 1:30. It is very important to follow the manufacturers instructions regarding the use of chilled pipettes, tubes and DMEM as matrigel will gel very quickly. Good luck!

I use matrigel diluted in DMEM/F12, usually I add 500ul of matrigel for 50 ml of DMEM/F12, but check the notice but this volume can change because it depends on the manufacturer instructions and the matrigel lot, ...in my case with lot i'm using is 310 ul for 50ml DMEM/F12...

As Leila told you it depends on the manufacturer instructions. Check the data sheet for the recommended dilution. Also take in mind that you have to defrost the matrigel in ice, because if you defrost it at room temperature it will be impossible to pipette. I usually defrost it in the freezer inside a box of ice overnight.

We use matrigel diluted in DMEM. We add 10ml of cold DMEM to the 10ml matrigel vial for a 1:1 dilution. These can then be aliquoted and frozen. We then dilute the aliquots 1:15, again in cold DMEM, so that the final working ration is 1:30. It is very important to follow the manufacturers instructions regarding the use of chilled pipettes, tubes and DMEM as matrigel will gel very quickly. Good luck!

Thanks very much for your kind replies. Can you tell me where the recommended ratio of dilution is? I can not find it in manufacturer instructions. My Matrigel is from BD Inc. with cat. 356231, lot: 41494. Can you show me a link?

I was checking for myself so I found this from I guy working in a lab

Matrigel Coating of Flasks: 500μl Eppendorf of Stock Matrigel was taken out from the freezer and placed in fridge (40C) for about 3 hours to warm it gradulally. Matrigel is diluted using 50ml chilled knock out serum free DMEM. 250μl of matrigel was pipetted into the 50ml universal tube with 50ml knock out serum free DMEM. Then the rest of the eppendorf was washed with 0.5ml of media and added into the 50ml universal tube as well. The media was mixed rapidly and 3ml-4ml of matrigel solution w

as added quickly into each T25 flasks (4 flasks were prepared) before the matrigel sets. Flasks were left in the hood at least 1 hour to coat the flask surface.

but check online before use it

I think it depends what matrigel you are using. We have matrigel that we don't dilute.

You can find it in the second page (verso) and the top of the paper joined to your bottle and your certificat analysis paper...

Matrigel should be used in µg/ml. I dilute it to a concentration of about 80µg/ml (1mg in 12ml od ice-cold DMEM-F12). This works very good with hESC H9 line. Coat dishes for at least 1h @ +37°C. Use directly.

The 1:30 dilution is the protocol we use for GFR matrigel. For HESC qualified matrigel the datasheet sent with the product has batch specific dilutions.

To use it with muscle cells we dilute it at 10% v/v, usually we use 500ul in 5ml of plain DMEM. But as already said in other comments, it all depends from the manufacturer.

We are using Matrigel as well for a lot of our experiments, espessially for the feeder-free cultivation of hESCs. The most important part is really to keep the matrigel during the whole prepartion and pipetting cold because the quality in the end will then be much better. Ideally use cold tips and pipetts to aliqout the matrigel in an appropriate volume for your needs and store the small amounts for dilution at -80°C. We are using the hESC qualified Matrigel which is already pretested for hESC cultivation and there it is definetly written how much µl you should use for 25ml or 50ml in DMEM/F12 (ideally knockout Media which are normally used for ESC cultivation).

From my own experience the written Matrigel amount of BD (now it should be Corning) are more than enough to guaranty a good quality for hESC cultivation that you can test the batch as well in different concentrations. It is normally said that you should not store the Matrigel coated plates longer than 7 days at 4°C but as well there the timeframe is longer (up to 2 weeks was never a problem even sometime 3weeks).

Overall the aliquotation and the cold handling is the most important part to get a good matrigel layer on your dish. But the amount you need definitly depends on the batch and the quality - so maybe testing is adviseable.

I always used a 1:8 dilution.

Dilute the basement membrane extract (tradename Matrigel) with cold (on ice) DMEM. Day before the dilution transfer Matrigel from -80oC to fridge (+4oC) and keep it in the fridge overnight on ice (ice will melt on the next morning, so add the appropriate amount of it). The final concentration for dilution is 50 to 100 ug/ml. All dilutions must be performed on ice using pre-cooled glassware .

The best dilution media is DMEM (50%) + F12 (50%) mixture.

Matrigel (growth factor–reduced; BD Biosciences, 8.7 ug/cm2). Zhang et al. Circulation Research. 2012; 111: 1125-1136

We dilute Matrigel 1:100 with cold DMEM/F12, coat the well and incubate at 37 degrees C for 1 hour.

I usually use this kind of approach with the Matrigel coating for both hESC and hIPSCs, since Matrigel stock concentrations varies from lot to lot:

Firstly I use growth factor reduced Matrigel from BD and prepare aliquots of it such as I have 2mg Matrigel/24mL DMEM/F12 (w/ HEPES). With such concentration, I can plate 4 6-well plates with 1mL Matrigel/well (that gives you a basis for adjusting the amount of Matrigel you may need for your application). And I store my plate in the incubator at 37ºC for at least 1 hour (up to 5 days) and worked wonderfully well for H9, IMR90-c4 and some other particular iPSCs.

I have tried matrigel coating at different dilutions and with different medium, and what works best in my hands is 1:100 dilution in KnockOut DMEM, not DMEM/F12. As the others said, it is important to keep matrigel cold at all times, on ice, and pre cool the tips before using (matrigel gets rapidly gelified) After coating plates/dishes, I leave them under hood O/N with parafilm, and if you like to use them later, you could store them later at 4C for up to 3-4 weeks. Good luck.

Hi Dears

we are going to inject cancerous cell lines mixed with matrigel, the high concentration of Corning Matrigel Matrix (20 mg/ml) was uesd, for the first time we diluted the matrigel 1:1 with PBS, but no significant changes observe in the size of tumor.

Anybody know what is the best concentration of the matrigel for subcutaneously injection?

Dilution ratios are useless without knowing the protein concentration, which varies from lot to lot. I use 1 mg per 100mm plate, or that plate's equivalent, such as 3 60mm plates or 1 tissue cluster.

The OP asked about matrigel, not whatever it is you are selling this time. You have posted this exact same response in 14 threads so far today.