The yield of my LDHA protein is low and functional. As such, is there any way to increase the recombinant protein yield by manipulating the induction?

13 October 2017 1 357 Report

Regarding IPTG induction

There are many methods to optimize protein expression: time, time vs IPTG concentration, temperature, time of induction - cell induction in stationary phase or for same protein in logarithmic phase is better. You can also change expression strain and, aerobic or anaerobic conditions, using protease-deficient host strains and much more. I hope that the attached publication will be useful for you.

Badges
Science topic

More Hendra Goh's questions See All

What are the moving particles in the organotypic slice culture?

Hi everyone, I experienced some moving particles in my mouse hippocampal slices. The media looks clear. Any idea what these moving particles might be? Thanks you!

22 July 2024 3,734 0 View

Best book of rock mechanics, rock engineering, mining geotechnic?

What is the best book of rock mechanic, rock engineering, geotechnical engineering, and soil mechanics? . Iam a geothecnical mining engineering and I know that "Rock Slope Engineering" 4th and 5th...

13 July 2023 4,637 4 View

Suggestion of antioxidant/stabiliser for receptor medium in skin permeation study?

We are doing ex vivo skin permeation studies for an easily degraded drug. Any suggestion for antioxidant/stabiliser to minimise degradation of drug while not affecting the skin integrity?

11 September 2021 6,820 2 View

Why is Tris buffer added during microsome preparations?

Hi, I am planning to make microsomes and wondering what components I should be adding to my 0.25M sucrose. I am being mindful that different additives can influence my microsome activity, which is...

04 August 2021 4,961 3 View

Missing listwise in SPSS?

I ran correlation and t-test with n = 222. However, when I ran hierarchical multiple regression, the n came out eventually is 221. I saw a "missing listwise" word before the output table. Does it...

01 August 2021 6,722 5 View

I have 2 questions about human genetics?

1. Can the DNA mutations that do not affect the phenotype be used as the markers to develop human genetic maps? 2. Can the human genetic maps be developed based on DNA mutations that affect phenotype?

03 April 2021 4,403 3 View

HPLC separation for methyl salicylate, camphor, thymol and menthol?

I have been recently trying to develop a method to separate the above compounds. Most methods tried were adapted from literature using RP C8 or C18 column with ACN/MeOH and buffered water at pH 2...

28 December 2020 4,876 6 View

How is a 5-log reduction in CFU/ml not statistically significant?

I performed a two-tailed t test with Welch correction on a treated cell versus the untreated control. As shown in the figure attached, there is almost a 5-log reduction, but the t test gave a p...

28 September 2020 508 4 View

How is the quality of archived blood sample for HbA1c test?

HbA1c is an important indicator of long-term glycemic control with the ability to reflect the cumulative glycemic history of the preceding two to three months. Also, HbA1c is measured as the ratio...

16 September 2020 2,645 4 View

What is the function for all of the components in the GUS staining solution?

My GUS staining solution contained phosphate buffer, ferricyanide, ferrocyanide, triton X-100, EDTA, methanol and X-Gluc. Except for X-Gluc, anyone know the function or importance of other...

14 May 2020 557 3 View

Can I use a HisTRAP column for affinity chromatography?

I'm working on selecting antibodies against a recombinant protein that has a His-tag. My idea is to first bind the recombinant protein to a HisTRAP column and then use this column for an affinity...

07 August 2024 505 3 View

What is the best blank for nanodrop if I want to read a recombinant protein concentration?

Is it the "elution buffer" or the "dialysis buffer"? Note: I'll be using NanoDrop OneC

01 August 2024 967 3 View

How are aPTT and PT values reduced after hemodialysis compared to before dialysis value in patients with chronic kidney disease?

I am conducting my research regarding blood coagulation in patients with CKD. I observed a reduction in post hemodialysis aPTT and PT values compared to pre hemodialysis values. But most of...

30 July 2024 3,467 0 View

Problems with ion exchange chromatography?

I'm having problems with ion exchange chromatography. After applying my sample (a fluorescent protein), a preferential path inevitably forms. The matrix I use is DEAE-Sephacel. Could someone...

27 July 2024 6,605 0 View

How do regenerate Amberlite XAD 4 resin for further cycles of use for removing 0.5% Sarkosyl from recombinant protein solutions?

I use 0.5% sarkosyl to solubilise recombinant proteins contained in Inclusion bodies of E. coli. sarkosyl very effeciently solubilizes proteins from Inclusion bodies and I am trying to use...

25 July 2024 5,001 2 View

How can I calculate the SOD enzymatic activities using the spectrophotometer method?

I have to quantify the enzymatic activities of SOD in plants using a method that employs hypoxanthine-xanthine oxidase to create O2. The colorimetric reaction was performed by combining...

23 July 2024 8,492 3 View

How do nutrient transporters and aquaporins interact under abiotic stress in plants?

"The Correlation between Nutrition and Transport Mechanism under Abiotic Stress in Plants: A Comprehensive Review" by Muhammad Saleem, Jianhua Zhang, Muhammad Qasim, Rashid Iqbal, and Li Song,...

23 July 2024 9,109 0 View

Consequences of excess bases in the MCS after digestion?

Good morning, I am trying to produce a recombinant protein. Unfortunately I one of the two restriction enzymes that I use do not cut at the end of the MCS. After the digestion 15 nucleotides of...

17 July 2024 2,787 4 View

Unexpected 35 kDa Band Observed During Expression and Purification of Dengue 3 NS5 RdRp Domain ?

Background: I am developing an RdRp assay for Dengue virus-3 and need to express and isolate the NS5 protein's RdRp domain (~101 kDa). My expression vector includes a GST tag followed by a 6x His...

13 July 2024 9,950 3 View

THP-1 cells completely dying with PMA, help?

Have been trying to induce THP-1 using PMA at different concentrations (range from 5 to 50 ng/mL) but after 48h all the conditions except the control are mostly dead, checked by trypan blue and...

03 July 2024 3,884 5 View