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Questions related from Suipwksiow Wjkp
The antibody is against human protein, but with mouse IgG (as it is generated in mouse). Before totally humanizing it (which takes a lot of money and time), I want to validate its function in...
21 April 2022 9,955 0 View
Hi Everyone, I'm exploring the anti-tumor effect of some drugs. I'd like to test some FDA-approved drugs which are not used to treat cancer. My question is, if there is any anti-tumor effect for...
15 December 2020 1,653 1 View
Hi Everyone, I did numerous human T cell CFSE proliferation assays, but the peaks never shift. I put CD3/CD28 and IL2, and I can see that T cells did proliferate by my eyes (as there was increase...
09 December 2020 9,259 3 View
Hi all, I used tumor dissociation kit(Miltenyi Biotec) to first isolate cells from tumors. Then I washed cells twice with PBS, and cultured the cells in complete RPMI (5%FBS, and pen strep)....
05 November 2020 5,653 1 View
Hi everyone, The protein I studied is a membrane protein. Some study indicates that there is possibility that this protein can form a dimer. I did standard western blot for this protein and my...
21 October 2020 7,183 7 View
Hi all, My hypothesis is that CD99 binds to CD88 (two membrane proteins). My colleagues suggested me to do co-IP, but I don't have good western blot antibody against one of the protein. However,...
15 October 2020 9,404 6 View
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...
07 October 2019 7,407 3 View
I'm synthesizing a chimeric antigen receptor (CAR), and I have the whole sequence of the amino acid. Now the problem is, to ask company to synthesize the DNA of the CAR, should I send real gene...
07 February 2019 1,668 1 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...
04 January 2019 1,901 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...
30 November 2018 6,481 3 View
There are so many HDAC family member (1-11), but which is the one that is inhibited by HDAC inhibitor?? I ask this question because I found some of them seemed to be tumor suppressor, while some...
26 March 2018 679 2 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??
14 March 2018 4,638 3 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...
09 March 2018 5,172 1 View
I have the sgRNA vector from addgene(#75112). It has the BsmBI site. I've designed the sgRNA already. The problem is, should I also design a control sgRNA??? It seems the vector itself, (also a...
09 March 2018 3,764 3 View
Is there anyway to preserve human blood sample/PBMC for future isolation of lymphocytes? If not, or if the way will largely reduce the yield of lymphocytes and causes some changes, what's the...
08 March 2018 9,664 3 View
Is it possible that patients that got gene therapy may pass the engineered genes to their next generation? e.g, the virus gene therapy, deliver some genes to human body. Will the human pass some...
24 February 2018 2,524 3 View
I know Lenti vector is usually used for making virus and doing transduction. But can I use it for transfection??? Thanks
22 February 2018 5,712 4 View
These immune killers cells can kill target cells by releasing cytotoxic granules such as perforin and granzymes. Why don't perforin/granzymes released by NK or T cells kill NK/T themselves?
22 February 2018 7,853 3 View
Can you list some synthetic transcription factors that are common and effective? i.e DNA binding domain(recognize and bind certain DNA) + transcription activating domain(drive transcription)
22 February 2018 1,236 1 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...
15 February 2018 9,701 2 View
I've designed a recombinant protein made of 5 parts from 5 different proteins. And I have the sequence of the whole recombinant protein I designed. It's around 2kb. Now the problem is where/how...
22 January 2018 9,668 3 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??
20 January 2018 5,240 6 View
I'd like to analyze the RNA level of TCGA data. However, there are two categories of the data One is harmonized data, under this, you can choose the category of "transcriptome Profiling", and...
04 January 2018 763 4 View
I'm doing differential expression analysis of TCGA Ovarian Cancer between tumor and normal. For transcriptome profiling, however, there is no data for normal (either blood normal or solid tissue...
04 January 2018 9,250 1 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...
28 December 2017 5,852 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...
15 December 2017 3,524 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....
20 November 2017 481 1 View
I used 26 gauge needle for subcutaneous injection of matrigel:cancer cells. However, when i pull out the needle, the matrigel solution also came out. How to prevent the leaking?
13 November 2017 5,659 3 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...
26 October 2017 5,507 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...
26 October 2017 2,399 3 View
In tumor microenvironment, stroma seems to be an important factor that makes tumor immunosupressive. Is there any other enzymes like hyaluronidase that is proved to dissolve stroma components?
26 October 2017 1,183 3 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...
23 October 2017 9,347 1 View
I've read many reviews and papers about the resistance of immune checkpoint blockade. Some said that the resistance is due to too much inhibitory receptors/ligands on cancer cells, which would...
23 October 2017 2,172 3 View
The cell line used is mouse fibroblast cell line. Retrovirus carring a human gene with an RFP tag is used for transduction. Then puro selection and RFP+ sorting. Cells transduced show RFP+ and...
11 October 2017 9,568 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...
25 September 2017 3,110 2 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.
22 September 2017 5,134 3 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...
18 September 2017 9,452 1 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?
18 September 2017 5,882 3 View
CAR-T is kind of common strategy. I also saw people use iNKT cell to express CAR. What's the benefit of using iNKT cell instead of T cell? It seems that CAR-T cell could be toxic to recipient...
18 September 2017 7,132 1 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...
08 September 2017 2,800 5 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?
01 September 2017 9,215 3 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...
07 August 2017 7,898 8 View
I'm analyzing TCGA data. I'm comparing a gene's expression between tumor and normal. However, it is possible that the patients get any treatment before, which affect the genes expression. How to...
20 July 2017 747 2 View
I need to let the cell express a cell membrane protein. I'm going to buy a lentivector containing the cDNA of this protein. However, there are many transcript variants for this protein, how do I...
13 July 2017 6,241 0 View
Why use special competent E.coli for gateway plasmid clone? What's the difference between the gateway competent E.coli with normal competent E.coli?
10 July 2017 8,157 2 View
If I have a protein coding direction, from right to left, is the promoter region on right side or left side??
10 July 2017 5,252 3 View
I need to compare a gene's expression between tumor site and matched normal tissue from TCGA database. I've tried using Firehose to search differential expression of the gene among different types...
10 July 2017 1,540 8 View
I have a list of genes ranked according to their expression. I want to know which gene encodes transcription factor. I cannot search one by one, because there are too many genes. Any suggestions...
27 June 2017 1,986 5 View
Mutant kras leads to constitutive activation of downstream factors, what if we directly delete Kras in the tumor tissue? Will there be tumor regression? Also, people don't target KRAS is because...
23 June 2017 4,678 4 View
Can a gene's regulatory region overlap with another gene? I'm looking for promoter region of a gene. If I take 1kb upstream of TSS, the sequence will overlap with an upstream gene. Should I take...
14 June 2017 2,802 6 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...
01 June 2017 3,211 4 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?
31 May 2017 6,355 4 View
Can people use oncogene-specific antibodies with ADCC effect to target cancer cells? The mutant product of the oncogene can be presented by MHC 1/2 on cancer cells. Can antibody recognize the...
31 May 2017 9,061 0 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
23 May 2017 4,859 3 View
Is there any way to control MHC 1/2 presentation? To let it present my desired peptide?
18 May 2017 5,652 3 View
Can Cas9/gRNA recognize SNP? What is the sensitivity of Cas9/gRNA? If the sensitivity is not good, is there any enzyme in Cas family can recognize SNP? or have higher sensitivity? Thanks!
11 May 2017 9,921 3 View
i've been doing bio research for many years. but in terms of physics and philosophy, i'm pretty confused why proteins function. e.g., why does endonuclease know to cut DNA? What is the force to...
25 April 2017 5,789 5 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...
06 April 2017 3,831 4 View
We all use anti-PD1 or anti-CTLA4 to release the brake of immune response. And we all know that CD3/28 activator can always help T cell expansion and activation. So, why don't we use anti-CD3/28...
01 January 1970 6,799 1 View
