What is the typical shape of iPSCs under microscope?

I am trying to make iPSCs from Keratinocytes (HaCaT). Being a beginner, it's hard for me to judge morphologically about the right clone under microscope. Can you please guide me by looking at the...

01 February 2019 8,876 5 View

Unexpected profile of TNF signalling after stimulation with TNF-alpha?

Hey! I used mouse fibrosarcoma cell line (L929) and treated with 1ng/ml rmTNF-alpha and extracted protein at different time points (0min, 15min, 30min, 1hr, 2hr, 5hr, 8hr, 12hr). Then I performed...

11 December 2018 9,120 3 View

Which one is better choice out of HEK293-hTLR5 and HEK293-hTLR5-CD14?

Hello, I want to confirm the specificity of my protein to human TLR5. For that, I am interested to buy the invivogen cell line but I came across two types of hTLR5 cell lines: 293-hTLR5 and...

11 December 2018 8,001 0 View

For Immunofluorescence, Which dye is suitable in secondary antibody if your cells are GFP positive?

My cells are transfected with GFP plasmid. I want to detect a particular cell surface marker by immunostaining. Can I use Alexa Fluor 488 dye containing secondary antibody? Will it overlap with...

08 September 2018 7,386 7 View

Protocol for Sigma Alkaline phosphatase (ALP) Kit (86R)?

I found an old ALP kit in my lab (once bought by seniors) with no protocol inside kit box (may be lost). I could not find the protocol on Sigma website. If someone has the protocol then can you...

08 September 2018 7,572 0 View

Why round shaped cells started having arms?

I converted differentiated cells into round shaped de-differentiated cells in F12/DMEM medium. As soon as, I transferred them to maintenance medium (TESR-E8 medium) they started changing their...

07 August 2018 1,948 2 View

Dose independent behavior of small molecules on cells?

I am treating my cells (MEF) with three different concentrations (1um, 10um and 20um) of various small molecules (M1, M2,.....,M10). Some of them show dose-independent behavior on cells but others...

06 July 2018 5,142 5 View

Dedifferentiation caused the detachment of the cells?

On the way to iPSCs. The targeted cells started to become circular (indication of dedifferentiation). Before I go for their confirmation as iPSCs they started to detach from the plate surface....

06 July 2018 4,775 1 View

How to combine result of excitation and emission radiations detected by microplate reader as one reading?

I did ROS assay using H2-DCFDA but could not find the function of detecting 2 wavelengths ( ex: 485, em: 520) at the same time as single reading in microplate reader so I got readings separately...

06 July 2018 8,876 6 View

How to separate 5793bp and 6320bp cut DNA fragments on gel?

It seems to be easy process but I can hardly cut them separately under UV light (long exposure to our body is hazardous). I used 1% agarose gel so far and ran for 2 hrs at 50V. If someone has...

05 June 2018 2,823 4 View

Waters 2707 HPLC autosampler problem?

I am using a 2707 waters HPLC device. When I try to inject a sample, it says missing plate or rack. I changed the needle and calibrated its position but I still get the same problem. I even get...

02 March 2021 1,408 1 View

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02 March 2021 3,060 3 View

Should we change media after1st 6hrs after plating HUVEC bought from INVITROGEN?it's not recomended by invitrogen.Will DMSO present in stock matter?

We have HUVEC bought from Invitrogen. Cat no -C-003-5c. while first plating the cells we have bought the protocol suggests not to centrifuge the cell with media to remove the DMSO. Should we...

02 March 2021 3,713 2 View

How to normalise data on OPM package from R Studio ?

Good afternoon, I recently used OmniLog from BIOLOG for my experimentations : I tested the metabolism of different strains on 2 types of plates. I have 16 strains of 3 different groups...

02 March 2021 3,584 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

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28 February 2021 9,936 2 View

Benefits of the 24 welled plate?

We have always used the 96 welled plate, the 48 welled plate but apparently there is much evidence on the 24 welled plate and above all its benefits.It states it is the same you would use on ELISA...

28 February 2021 1,149 1 View

Can bacterial suspension be pipetted directly onto petri dish and agar poured onto it?

Hi, recently I encountered a protocol for plating which was not the usual method I have been practicing (pour LB agar onto plate, allow it to solidify, pipette bacterial suspension onto plate,...

28 February 2021 7,292 3 View