Missle thank alot can you explain me again why ı should not divide by the volume? I also remember that is not necessary. Can you give me an example? How much of the unkown sample in volume must be pipeted into the 1ml Cuvet?
Submit your paper to J Biol Methods today!
Pangea's Content
There have been 97 items by Pangea (Search limited from 12-April 20)
By content type
See this member's
#163303 Bradford Assay Calculation! Help!
Posted by
on 11 December 2013 - 12:21 AM
in
General Lab Techniques
#161940 Bradford Assay Calculation! Help!
Posted by
on 31 October 2013 - 11:44 AM
in
General Lab Techniques
Dear All,
I have simple problem about Bradford Assay Calculation. I not designed this and would never do it in this way.
(BioRad) BSA Standard Curve
Stock Solution: 1.48 mg/ml
1: 5.92 ug/ml (from Stock 4 ul into 796 ul H2O + 200 ul Dye Reagenz ) OD595 : 0.434
2. 8.88 ug/ml (from Stock 6 ul into 794 ul H2O + 200 ul Dye Reagenz) OD595 : 0.657
3: 11.84 ug/ml (from Stock 8 ul into 792 ul H2O + 200 ul Dye Reagenz) OD595 : 0865
4: 14.8 ug/ml (from Stock 10 ulinto 790 ul H2O + 200 ul Dye Reagenz) OD595 : 0.989
y = 0.23x + 0.177
R2 = 0.99
Lets take 6 ul an unkown sample without dilution and a absorbence of OD595 0.295:
0.295 = 0.23 x +0.177 ----> (0.295-0.177)/0.23 = x ---> 0.513 mg/ml ATTENTION: We dived by 6 ul to obtain the true conc. : 0.513/6 = 0.085 mg/ml.
Can you confirm the calculation neglecting to way it was generated.?
And we do not have a sample 0 the blank is not in the graph.!
Must we dived by 6?
Can you send me a protocol?
#154198 Bacterial Media Formulation
Posted by
on 24 April 2013 - 10:33 PM
in
Protein Expression and Purification
BL21 DE3 or Origami ? But if you pass me all formulation what you it would be great. I think i will try auto-induction media with 2 mM MgSO4 or higher. Some Trace Elements. I know LB, TB, SB, M9 etc. but some how people have good media formulation. Actually, I get with 2xTB at 37 C and 0.5mM İPTG an OD600 of 14 in a shaking flask (100 ml/500ml). Any suggestions?
#153915 Bacterial Media Formulation
Posted by
on 18 April 2013 - 12:09 AM
in
Protein Expression and Purification
Dear All,
I would like to reach a OD600 greater than 50 in flask and 100 in Fermenter. Do someone knows a good media beside LB, TB or SB?
Thanks in advance
All the Best
I would like to reach a OD600 greater than 50 in flask and 100 in Fermenter. Do someone knows a good media beside LB, TB or SB?
Thanks in advance
All the Best
#152180 Production of GST tagged fusion proteins in BL21 Star cells
Posted by
on 13 March 2013 - 12:18 PM
in
Molecular Biology
One question to you construct: Do you have additional amino acids after cleavage of your protein from your gst?
#149918 Interferon Alpha
Posted by
on 08 February 2013 - 10:34 AM
in
Protein Expression and Purification
Hi Everyone,
I can not express interferon alpha in bl21 cells. Does anyone have an idea or protocol. Vector is pET9a.
THANKS in Advance
I can not express interferon alpha in bl21 cells. Does anyone have an idea or protocol. Vector is pET9a.
THANKS in Advance
#148128 Manual/ User guide for opticell from thermoscientific
Posted by
on 15 January 2013 - 12:16 PM
in
Tissue and Cell Culture
#148092 western blot troubleshooting
Posted by
on 15 January 2013 - 08:26 AM
in
Molecular Biology
Welcome.
as almost says, we need more detail and specification of your problem. But it seems that you just have different amount of actin as a internal control. Thats because your loading conc. are different when you perform SDS-PAGE.You can calculate the amount maybe with densitometric methods.
as almost says, we need more detail and specification of your problem. But it seems that you just have different amount of actin as a internal control. Thats because your loading conc. are different when you perform SDS-PAGE.You can calculate the amount maybe with densitometric methods.
#148091 protein sample floating out of the tricine gel
Posted by
on 15 January 2013 - 08:19 AM
in
Protein and Proteomics
Check Loading Buffer again! Bromophenol Blue!
#148006 Alternative to pET Vectorsystem
Posted by
on 14 January 2013 - 08:28 AM
in
Molecular Cloning
There is no big problem with pET. Just in case.
#147980 Difficult Ligation - Details Inside
Posted by
on 13 January 2013 - 01:35 PM
in
Molecular Cloning
I guess you use Calcium Chlorid and using all tranformed E. coli to plated on Amp plates.
#147977 Difficult Ligation - Details Inside
Posted by
on 13 January 2013 - 01:28 PM
in
Molecular Cloning
Better to increase insert. On your Gel you dont see your expected size? Maybe difficult on this size of an insert. With recutting i mean where do you no that its not working?
#147974 Difficult Ligation - Details Inside
Posted by
on 13 January 2013 - 12:41 PM
in
Molecular Cloning
I already changes my mistake. Sorry.
#147972 Difficult Ligation - Details Inside
Posted by
on 13 January 2013 - 12:33 PM
in
Molecular Cloning
XbaI is dam sensitive. Spe1 is compatible with Xba1. Maybe, use klenow and ligate blunt end. Did you do Re-cutting? PCR with Taq or Pfu? Generally, try different ratio for ligation.
#147963 Stem Cell Donation
Posted by
on 13 January 2013 - 09:40 AM
in
Stem Cell
Cheers on both.
#147962 pET 32 (+)
Posted by
on 13 January 2013 - 09:38 AM
in
Molecular Cloning
What means the "+" in the vector description? What kind of orientation is it?
#147961 Alternative to pET Vectorsystem
Posted by
on 13 January 2013 - 09:35 AM
in
Molecular Cloning
Any suggestion about other vectorsystem than pET?
#147958 High Yield Protein Expression 10g/L
Posted by
on 13 January 2013 - 09:30 AM
in
Protein Expression and Purification
So you mean replacement by centrifuging the cells and renewing the media. Can you give me more details like an protocol?
#147957 Stem Cell Donation
Posted by
on 13 January 2013 - 09:28 AM
in
Stem Cell
Germany
#147950 isolating DNA from cells in suspension
Posted by
on 13 January 2013 - 07:51 AM
in
Molecular Biology
Getting rid of unnecessary molecules in media. And handling with less solution (concentrating).
#147949 Software for scanning of specific aa sequence in proteins
Posted by
on 13 January 2013 - 07:45 AM
in
Bioinformatics and Biostatistics
#147948 Stem Cell Donation
Posted by
on 13 January 2013 - 07:40 AM
in
Stem Cell
Where can I type me and donate stem cells?
#147947 Purifying a "protein"
Posted by
on 13 January 2013 - 07:12 AM
in
Biochemistry
Run a SDS-PAGE and do Silver-Staining. Measurement on 280 nm for Purity.
#147946 Suggestion for a book!
Posted by
on 13 January 2013 - 06:49 AM
in
Immunology
Listen type into google search as follows: Cytokine filetype:pdf
The part filetype will give you just PDF files and gives you a smile in your face:). You can do this also with doc, xls, etc. Enjoy the discovery.
Books:
Cytokine Protocols (Methods in Molecular Biology)
http://elib.fk.uwks....ogy Series].pdf
http://www.ctf.edu.t...1_Cytokines.pdf
The part filetype will give you just PDF files and gives you a smile in your face:). You can do this also with doc, xls, etc. Enjoy the discovery.
Books:
Cytokine Protocols (Methods in Molecular Biology)
http://elib.fk.uwks....ogy Series].pdf
http://www.ctf.edu.t...1_Cytokines.pdf
#147945 How to raise polyclonal antibodies against whole bacteria live &/ or inactiv
Posted by
on 13 January 2013 - 06:41 AM
in
Immunology
Thats just general information, but which bacteria you want to produce rabbit Ab. It will be take 3 month i guess in rabbit. Whole bacteria means just its capsule, isnt? You might look up the consistence of the capsule. It dosent matter if it Gram-negative or -positive, isnt? But i guess it will be unspecific Antibody. Do your Bacteria has specific marker molecules on his surface?
http://www.uniprot.org/keywords/875
http://ecvam.jrc.ec....hopReport35.pdf
http://www.uniprot.org/keywords/875
http://ecvam.jrc.ec....hopReport35.pdf
