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Questions related from Suipwksiow Wjkp
I'm recently buying some anti-CD3 and anti-CD28 to activate T cells. I also notice some of this clones have been conjugated with fluorescence for the purpose of labeling T cells for flow...
10 October 2019 1,068 4 View
I have many single-cell sequencing data. I checked my gene of interest in these data. However, all of these single-cell sequencing data showed that my gene has no expression, no matter in immune...
01 January 2019 3,783 3 View
I was doing an over-expression experiment of a gene for a year. It is a surface protein. The biggest trouble is that I can never detect the gene overexpression by western blot. Interestingly, when...
11 November 2018 7,299 3 View
When I first learn cancer biology, my teacher told me that cancer is usually caused by the loss of tumor suppressor , which is like you loss your brake. After it losses the suppressor, it tends...
06 June 2018 8,164 2 View
Are there any mouse cancer cell lines that can form tumor in nude mice but not in normal (immune competent) mice? Please give me some examples. Thanks.
05 May 2018 6,779 2 View
AI seems to be pretty popular these days and there are many applications in our daily life. But are there any AI designed to help our researchers to solve any biology questions?
05 May 2018 4,130 3 View
According to the website https://www.addgene.org/14883/, it has bleomycin resistance. However, bleomycin can be used both for bacteria selection and cell selection. Then what is the cell...
04 April 2018 3,897 1 View
The two ends of my gene have same restriction sites. And the destination vector also has two same sites. Can I clone the gene directly to the destination vector? I know it's possible that the gene...
04 April 2018 8,421 4 View
Why don't use ampicillin to select transduced mammalian cells? also, I saw some lenti-vector is Bleo Resistant, is bleo used as bacteria selector or mammalian cells selector?
04 April 2018 7,750 0 View
Can I use NEB mutagenesis -- substitution method to clone sgRNA into the vector? I felt it's much easier. Just design the primers containing the sgRNA bp, conduct PCR and do a ligation. And most...
03 March 2018 654 1 View
For CRISPR, transfection or transduction? The CRISPR vectors I bought is lenti vector, so I can either do transfection or transduction. But which is better??
03 March 2018 2,293 4 View
Recently check the cell lines, 60% of the cell lines are infected with Mycoplasma. Some common antibiotics for treating Mycoplasma were used to treat cells for 3 weeks, but mycoplasma is still...
02 February 2018 4,036 3 View
I'd like to analyze the data in patient samples. I tried searching in GEO database by typing in some key words, however, no relevant studies come out. Is there any other database that I can find...
02 February 2018 7,663 2 View
Is there any transcription factor that has two parts and only function when both parts are expressed? one example is antibody, that has heavy chain and light chain and can only function when...
02 February 2018 3,889 3 View
I know Lenti vector is usually used for making virus and doing transduction. But can I use it for transfection??? Thanks
02 February 2018 6,474 3 View
Recently I somehow overgrew the bacteria in LB and get a low yield when extracting DNA. To get the highest yield, how many hours is should I grow bacteria? And how can I know when the bacteria...
01 January 2018 8,497 5 View
Theoretically, if the promoter region of a gene is highly methylated (hypermethylated), the transcription level should be low. However, I notice that in many cases, when the RNA level of a gene...
01 January 2018 4,718 5 View
The growth instruction for a CRISPR plasmid said that the plasmid need RecA negative E.coli strain like Stbl3 to grow. What is special for RecA negative strain? Why the clone need RecA negative??
01 January 2018 7,591 7 View
I have CHO cells that transduced with two vectors expressing light chain and heavy chain of an antibody respectively. How do they automatically associate with each other and form antibody? What...
12 December 2017 1,578 4 View
Among anti-CD3, CD28, OX40, 41BB, which is better to be bound antibody and which is better to be secreted antibody in order to stimulate T cells? And why there is such a difference of function...
12 December 2017 5,093 1 View
cut part of the tumor sample into a cassette, for IHC use; extract RNA from it, to run rt-qpcr, for mRNA analysis ; extract protein from it, for western blot; tumor dissociation, analyze immune...
11 November 2017 1,415 1 View
When I treat mice with two immune checkpoint blockades (release two breaks at the same time), it looks that the combination treatment is very toxic to mice, which seems due to the cytokine storm....
11 November 2017 1,305 1 View
There is orthotopic injection of 4T1 into mammary fat pad, there is also subcutaneous injection to the mammary region. I also see people use 4T1-luc for i.v tail injection(I don't have 4T1-luc...
10 October 2017 6,660 1 View
There are so many researchers developing inhibitors for tumor. For those tumor that has a single and specific shape, what is the main treatment? Isn't the tumor removal surgery much more faster...
10 October 2017 8,694 4 View
There are many problems with drug infiltration when using i.p./i.v. injection, because tumor has ECM outside and less vascular inside. For drugs that can directly function on tumor cells instead...
10 October 2017 2,168 2 View
For antibody treatment, the antibody always has sequence that the body never meet. So is it possible that the body can regard the antibody as a foreign antigen and delete it????
10 October 2017 1,212 3 View
I have this question for a long time. My boss told me every time I thaw a tube I should let it totally become liquid. What if I use the incompletely thawed liquid? Will the concentration be higher...
10 October 2017 7,264 2 View
It looks that some of my cell lines show two bands for vinculin on WB. The two bands are very close to each other, and they are on the right MW. I washed the cells with PBS for several times, so...
09 September 2017 4,433 2 View
How to design the primer for Sanger sequencing? I'd like to know the sequence of a 2kb inserts. Is there anything special that should be noticed? From primer to the interested sequence, how many...
09 September 2017 2,020 6 View
I've never done Luminex assay but only ELISA. Why do some people use Luminex assay but not ELISA to detect cytokines in supernatent? What is the advantages of Luminex assay?
09 September 2017 8,392 3 View
I'd like to fuse 3 fragments into a vector, but they have RE sites. How to connect them together without adding extra space? I don't wanna additional a.a to be translated.
09 September 2017 2,250 3 View
I'm overexpressing a gene in a fibroblast cell line. I also over-express it via transfection in 293T as a positive control. After stably transducing the cell line, I detect the gene expression by...
09 September 2017 6,725 2 View
I want to study a immunosuppressive gene. I knock it out in a cancer cell line and transplant it into B6, and compare its growth with WT tumor. I would expect that KO tumor grow slower than the...
09 September 2017 3,778 1 View
I'd like to do a lenti-virus transduction as soon as possible. Is it OK to thaw cells , directly plate them into 6-well plate, and after overnight, do the transduction?
09 September 2017 2,218 3 View
I'd like to get the sequence from an antibody because I want the variable region of the antibody to get expressed in my cells. 1. How to get the sequence of the Ab? 2. After knowing the sequence,...
09 September 2017 1,736 7 View
Is there any book that systematically introduce how to make genetic design? Like introducing various promoters, elements (such as P2A, IRES, etc), CAR-T design, etc?
08 August 2017 9,441 1 View
I first digested two vectors (the vector containing my interested gene and the destination vector). I ran on gel, cut the band, and did gel extraction. Then I incubate the inserts with the...
08 August 2017 1,014 9 View
I found a lot of commercial plasmids are bidirectional, that they have genes in opposite direction. How does that work? PS: See the attached, the arrows point to the opposite direction.
08 August 2017 8,081 9 View
I have a ORF library. I'd like to have one gene on one cell. How to do the transfection/transduction?
07 July 2017 407 2 View
How to find desired cell lines of certain mutations and certain types? For example, I'd like to study PTEN in prostate cancer. I need PTEN deficient cells and PTEN wild type cells. For each kind...
07 July 2017 7,596 5 View
The ligand of a cell surface receptor is unknown. I'd like to figure out what it is. Is there any good assay to identify new ligand for a cell surface receptor? Any good ligand/receptor library to...
07 July 2017 660 1 View
The first step of ELISA is to coat plate with antibody (sandwich ELISA) or antigen(ELISA). How does the antibody or the antigen bind to the plastic plate? According to some website, it is said...
06 June 2017 7,771 26 View
It is known that immunogen should be foreign to the host. e.g. if the immunogen(the peptide) is from mouse, then the host should be rabbit. However, if I use rabbit as host to produce antibody...
06 June 2017 4,847 5 View
We always buy mouse invivo antibody from Xcell, but rarely see other company selling the antibody against the same protein. I also see some people already applied the patent for the antibody use....
06 June 2017 8,971 2 View
Why do some antibody have blocking effect while some have agonist effect? For a co-stimulatory receptor/ligand, an antibody against it is usually agonist. Can an antibody against it have blocking...
06 June 2017 7,507 3 View
Some mAbs can only be used in WB, IHC, while some mAbs can be applied in vivo. What is the difference between these mAbs?
06 June 2017 9,042 4 View
We usually have a target gene, and search it for survival rate with or without the gene alteration. However, is there any way to do the opposite search? i.e., from two group of patients separated...
06 June 2017 6,733 1 View
Is there any way to control MHC 1/2 presentation? To let it present my desired peptide?
05 May 2017 5,021 3 View
It is known that 293T constitutively express SV40 large T antigen, which can bind to SV40 enhancers. Does that mean that SV40 promoter is the best promoter in 293T cell?
05 May 2017 7,821 3 View
Is EMT cancer-type-specific? Or does EMT occur in all kinds of tumor? I'm curious how specific the EMT signature is among different types of cancer. Is there any cancer type that EMT prefer? Thx
05 May 2017 2,319 4 View
I have a strain of bacteria. it's just a common bacteria without any genetic engineering or other processing. The instruction said that it can grow in normal LB broth. But I just worried if there...
04 April 2017 2,453 5 View
I'm growing a yeast-like fungus in a culture medium. After several days, I remove cells via filtration and get supernatant. Then I need to sterilize the supernatant to make sure it is bio-safe so...
04 April 2017 7,237 21 View
