Search found 259 matches

by biology_06er
Sun Jun 24, 2012 11:14 am
Forum: General Discussion
Topic: Flow cytometry-antibody conc.
Replies: 1
Views: 1902

Problem solved!
by biology_06er
Sun Jun 24, 2012 11:13 am
Forum: General Discussion
Topic: washing bacteria
Replies: 1
Views: 1658

washing bacteria

Hi there, What does it mean when it says "wash twice with PBS" regarding bacteria..does it mean after I spin down my bacteria, I pour off the supernatant and then add PBS and then spin again, pour off s/n add more PBS and spin... OR DOES IT MEAN just add PBS ensuring the pellet doesn't bre...
by biology_06er
Sat Jun 23, 2012 4:22 am
Forum: General Discussion
Topic: Flow cytometry-antibody conc.
Replies: 1
Views: 1902

Flow cytometry-antibody conc.

Hi there, I'll be doing FC for the first time next week, and I have a protocol but I'm slightly confused about one thing...after I resuspend my cells in PBS/BSA it tells me to take 200uL of cells and incubate with 1:200 of primary antibody...so does this mean if I have 200uL of cells, I add 1uL of m...
by biology_06er
Sun Jun 17, 2012 4:40 am
Forum: General Discussion
Topic: pET32a vector size limitations
Replies: 0
Views: 1549

pET32a vector size limitations

Hi there,

Anyone know about the size limitations for the pET32a vector...I've read on a few posts thats proteins with a MW of >70 kDa don't express but haven't been able to find any solid references (ones that I can actually cite?)

Anyone know any good articles?/refs stating this?

cheers,
b_06er
by biology_06er
Sat Jun 16, 2012 12:41 am
Forum: General Discussion
Topic: Restrction digest on a plasmid--how many fragments?
Replies: 1
Views: 3372

Restrction digest on a plasmid--how many fragments?

Hi there, Soo, cloning for about a whole year I kinda just thought about this question!! ....If I have a vector (pBS) for example...and my gene of interest that I wish to clone into it and then GOI has a EcoRI site at N-terminal and BamHI at C-terminal...so I would cut my GOI with those enzymes, and...
by biology_06er
Wed Jun 06, 2012 12:13 pm
Forum: Microbiology
Topic: adherence to host cell-capsule hindrance
Replies: 1
Views: 5861

adherence to host cell-capsule hindrance

Question, and sorry if it sounds dumb!

If a bacterium is covered by a capsule, and you have a possible adhesin embedded in the cell membrane, does the capsule not hinder the ability of the adhesin to adhere to the host cell?
by biology_06er
Wed Jun 06, 2012 2:38 am
Forum: General Discussion
Topic: What stats test should I use?
Replies: 1
Views: 2090

Re: What stats test should I use?

anyone? I tried doing the RM anova but it came out as sig..but clearly I had no binding..soo what am I doing wrong..essentially the -ve control values were the same as by test readings..the curve shows a very very slighly increase from lowest conc. to highest conc. If i wanted to test if there was a...
by biology_06er
Sun May 27, 2012 3:56 am
Forum: General Discussion
Topic: What stats test should I use?
Replies: 1
Views: 2090

What stats test should I use?

Hi there, I recently did an ELISA and have no idea what type of stats test to use?...I haven't done a single stats paper/stats related anything in over 6years..so am struggling to analyze my current results. I basically did the standard elisa,,,,adding various amounts of protein to coated wells...an...
by biology_06er
Fri May 25, 2012 4:37 am
Forum: General Discussion
Topic: E-cadherin/calcium
Replies: 1
Views: 2273

E-cadherin/calcium

Hi there, I recently did an ELISA with E-cadherin plated onto a 96-well plate then protein then primary anti and then finally secondary antibody. After I had finished it, I thought about the fact that when I plated my protein I had it in PBS with NO calcium. My e-cadherin was plated in PBS/Mg/Ca o/n...
by biology_06er
Sun Apr 29, 2012 1:15 am
Forum: General Discussion
Topic: reconstituting protein
Replies: 3
Views: 5411

Re: reconstituting protein

Hi, thanks for that...so another question following on from it..if I have 100ug/mL of protein and I plan to run an ELISA using it..and to each well I need (1.5 ug/mL with 100 uL/well) So for example, if I wanted to use 50 wells with 100uL in each well and 1.5ug/mL in each well would I use the formul...
by biology_06er
Wed Apr 25, 2012 10:33 am
Forum: General Discussion
Topic: reconstituting protein
Replies: 3
Views: 5411

reconstituting protein

Hi there, I was sent a protein (100ug) and the data sheet tells me to reconstitute at 100ug/mL in PBS..does this mean I just add 1mL of PBS to the powder form I have hence I'll have a final conc of 100ug/mL... what if I had a powder form of 50ug (as you can get this conc as well sent and the data sh...
by biology_06er
Mon Apr 09, 2012 4:37 am
Forum: General Discussion
Topic: ELISA modification
Replies: 1
Views: 2664

ELISA modification

Active Members 34 posts Posted Yesterday, 10:12 PM Hi there, I've done a number of ELISAs previously with the below protocol: 1:Coat plate with 1ug/ml protein 2:Leave at 4C o/n 3:wash 4:block/incubate 5:wash 6:add antibody/incuabte for 3hrs 7:wash 8:add secondary antibody/incubate for one hour 9:was...