Does anyone know any good anti-fliD (HAP2) antibodies?

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Why my Gibson assembly doesn't work?

I'm assembling 3 PCR fragments with my vector pBAD and transforming them into dh5a. The second PCR fragment has 4 variations to it and the template for this PCR fragment comes from a plasmid. I've...

01 February 2019 5,780 3 View

Does anybody know where to purchase anti-H7 for flagellin in escherichia coli?

Im specifically looking for an anti-mouse antibody for flagellin (antisera H7). I've checked papers and googled extensively but couldnt find it. Any ideas?

03 April 2017 2,233 2 View

Does anyone have a good protocol for flagella preparation/purification?

I need a protocol to shear flagella off E.coli O157:H7. I have been using a protocol online that is basically resuspending an overnight culture in a syringe and then, spinning it down to acquire...

02 March 2017 6,724 2 View

How does one crystallise a protein with the presence of TCEP as the reducing agent, and with it's co-factors (NAD+ and Fe2+)?

I have a protein which I know is stable with the presence of co-factors (NAD+ and Fe2+). Recently i have been trying to purify it with the TCEP as a reducing agent since my protein is known to...

10 November 2016 289 2 View

Why mbp-cleaved protein disappears after anionic exchange?

I am purifying a protein (47 kDa) that is fused onto an MBP (42 kDa). The PIs for both proteins are 6.48 and 4.9 respectively. After purifying it using a dextrose column, I have buckets of fused...

04 May 2016 9,084 8 View

How to find self-associating motifs in a protein sequence?

Hello, I have a protein that self associates into multimers and I have confirmed that using AUC. I would like to find the seqeuence in the protein that is responsible for self-association. Does...

04 May 2016 3,271 3 View

Why did the cleaving of my MBP tag on my protein failed?

I'm purifying a protein that is fused onto an MBP tag (42.5 kDa). My protein size is roughly the similar size of the tag (47kDa). Together, they form a complex about 89 kDa. After a run through a...

03 April 2016 8,736 3 View

How do I know if a flagella is in R or L conformation?

I have sequenced my sample and found some mutaions in the fliC gene. I have found that they have introduced some aa changes however, I would like to know if this changes the conformation into R or...

02 March 2016 7,551 1 View

How to generate GNOM file (.out) for DAMMIF runs?

I have .dat files from scatter that contain the Dmax of a protein and now I want to generate ab initio models. however I dont know how to generate .out files that are needed byy DAMMIF programs on...

09 October 2015 9,188 1 View

Why does the pressure alarm keeps going off during Nickel column purification?

I'm using a new column (5 ml Ni NTA HiFliQ column from Generon) to purify my protein AdhE which is Histagged and sized 96 kDa. However, during purification, the alarm pressure keeps going off even...

07 August 2015 5,258 10 View

Mouse CD3 antibody sequence?

Can anyone share any reference that gives mouse specific CD3 antibody sequence. I have been searching for this since few days and no luck. Thank you very much

08 August 2024 9,157 4 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

Do I need specific antibodies to stain for a fusion protein?

I am staining some brain sections stored in cryoprotectant that express a Histone H2B- GFP fusion protein that were generated ~10 years ago. I know I need to enhance signal with an anti-GFP...

07 August 2024 5,338 2 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 to make a Lateral flow control line more uniform?

We have a lateral flow test on Sartorius 140 NC. The conjugate is gold-monoclonal antibody. Using different control lines (GAM, protein A, protein G) we get a very strong leading edge with reduced...

31 July 2024 4,510 3 View

Hello researchers, how can one get the optical density of a filamentous algae?

i am comparing the growth performances of filamentous and non filamentous forms. in a case where I can not count the cells of the filamentous algae using a hemocytometer, is it okay to do a...

30 July 2024 5,985 2 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

Why my IgG antibody looks fragmented on non-reducing SDS-PAGE?

I am performing IgG purification and I have to show my results on SDS-PAGE. I use 10% tris glycine gel and prepare the samples under non reducing conditions. I am new to antibodies and therefore...

23 July 2024 6,664 6 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