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Questions related from Anita Ma
I found the phosphorylation level of ERK1/2 in my cell sample increased when I treated the cell with a medicine, but the mRNA level of my target protein decreased obviously. Is that normal? Coz...
03 February 2025 3,739 4 View
As we know that Actinomycetes produce most of the clinical used antibiotics, therefore, what mechanisms do they use to protect themselves from the antibiotics they made?
07 December 2024 621 3 View
As we know that antibodies are the products of an antigen invasion during humoral immune response. So where does the anti-B antibody come from for a type A blood person when no transfusion occurs?
20 September 2024 6,106 2 View
Trouble 1: I ran a PCR reaction and found a low molecular weight band (100 bp) when doing electrophoresis. 1ng plasmid (OD260/280=1.84) was used as template. Final concentration of primers was...
14 June 2024 5,611 6 View
I want to detect my target protein through WB and it worked well before. But now I could not get any signal for both my target and the GAPDH, so I ran SDS-PAGE. I expected many bands on the gel of...
24 May 2024 683 6 View
I want to copy a target gene from cDNA into a plasmid. The primers were designed according to the CDS sequence from NCBI. But when I performed PCR reactions I could not get any target bands. So...
16 April 2024 2,170 4 View
I found to use a wrong secondary Ab for my WB when I visualized it with ECL, can I wash with TBST for several times and incubate the right secondary Ab? Another question is why my bands are not...
17 February 2024 3,237 4 View
When I overexpressed a protein in a cell line, I found decrease of phospho-PLC (phospholipase C) at the protein level through WB. But when I knocked out the protein, there was no difference of...
11 January 2024 8,258 3 View
I want to use the Cre LoxP to knock out my target gene in a mammalian cell line. According to the mechanism, I need to add two loxP sequences at both ends of the gene through homologous...
22 November 2023 5,203 1 View
After proteomics analysis of my samples, a specific phosphorylation site on a kinase was found. But when I want to validate the result I could not find any commercial ab to that phosphorylated...
15 November 2023 436 6 View
This is my PCR 1.2% agarose gel result. I used a 25uL reaction system (0.3uM primers, 12.5uL Taq master mix, 1uL genome template and 5.5uL H2O). The primer Tm is 62℃, and reaction procedure is 94℃...
15 August 2023 1,409 5 View
I want to find out potential mechanisms of the effects of a certain gene on recombinant protein productivity . So proteomics analysis was performed between a high expression of my testing gene and...
28 March 2022 2,962 2 View
I am doing over expression of an endogenous target gene in a CHO cell line. When I did PCR (whole length of the gene) to verify the existence of my target gene, I could only see the gel band when...
28 October 2021 6,508 6 View
After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...
26 February 2021 6,172 3 View
