Could I trust in qubit for protein quantification?

I need quantify the protein amout from human DCs lysed by RIPA buffer ( 150 mM NaCl, 1% NP-40 or Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS, 50 mM Tris, pH 8.0 ) to use in a immunoblot....

11 December 2019 3,701 3 View

Is it possible to use the same permeabilizing agent to perform intracellular staining of cytoplasmatic and nuclear proteins in flow cytometry?

I work with human mo-DCs and I need to detect cytoplasmatic and nuclear proteins by flow cytometry using flurochrome-conjugated antibodies (direct detection), that is, I need to permeabilize both...

04 May 2019 4,991 5 View

What is recommended temperature to perform fixation with formaldeihyde or paraformalde to intracellular staining by flow cytometry of culture cells??

04 May 2019 3,390 4 View

There is a need to fixation step to detect soluble proteins in cytoplasma or nucleus by flow cytometry?

I need to detect and measure the abundance of cytoplasmatic proteins (AIM2, GBP1, GBP4) and nucleic proteins (IFI16), all apparently soluble, using direct detection by flow cytometry with...

04 May 2019 5,624 3 View

Why mercaptoethanol in THP-1 culture medium?

I have purchased THP-1 cell line from ATCC and they recommed mercaptoethanol. It is a reducing agent, what is the function of this kind of agent in culture medium of THP-1 cells? How...

04 May 2019 9,930 3 View

What is the best permeabilizing agent to detect soluble cytoplasmatic and nucleic proteins by flow cytometry?

I need to detect and measure the abundance of cytoplasmatic proteins (AIM2, GBP1, GBP4) and some nucleic proteins (IFI16)using direct detection by flow cytometry with fluorochrome-conjugated...

04 May 2019 7,582 2 View

What is antibody concentration recommended to western blot?

I need to detect and measure the abundance of cytoplasmatic proteins (CASP1 an GSDMD) by western blot in human mo-DCs, and some companies where I purchased the antibodies do not specify a...

04 May 2019 1,385 5 View

What is the difference between formaldehyde and paraformaldehyde in cells fixation for intracellular staining using flow citometry?

I need to detect and measure the abundance of cytoplasmatic proteins (AIM2, IFI16, GBP1, GBP4) using direct detection by flow cytometry with fluorochrome-conjugated antibodies in human mo-DCs and...

04 May 2019 6,491 4 View

How many human primary cells I need to quantify protein abundace in western blot analysis?

Someone that work with human primary cells and western blot: how many cells I need to identify protein abundance in western blot analysis? I want to quantify Caspase-1 and gasdermin D

31 December 2018 5,105 5 View

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I want to detect caspase 1 activation in human monocytes-derived dendritic cells after interaction with my pathogen.

10 November 2018 2,081 4 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

Buffer to disrupt platelet alpha granules for PF4 quantification?

Hello! I want to quantify by ELISA the secreted (from platelets poor plasma) and the non-secreted (from platelet lysate) PF4 before and after TRAP stimulation. I will use the ELISA from R&D...

03 March 2021 1,499 2 View

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02 March 2021 3,060 3 View

Sample dilution for quantitative analysis using ELISA?

If the detection range is in ng/ml but the reference range is in ug/ml for a molecule or protein in serum or plasma .how to dilute and what is the initial volume to be taken for quantitative analysis

02 March 2021 7,670 3 View

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

Does anyone use antinuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples?

I am looking for anti-nuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples. Any suggestions highly appreciated. Thank you.

28 February 2021 6,512 3 View

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28 February 2021 9,936 2 View

Anyone know of a good progesterone antibody?

I am looking for a good progesterone antibody to quantify levels of progesterone in zebrafish. Applications would be ELISA, and immunohistochemistry.

28 February 2021 5,929 2 View