User:Wilfred J. Poppinga/Notebook/cAMP compartmentalization/2010/03/16: Difference between revisions
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==Materials & Methods== | ==Materials & Methods== | ||
===Materials=== | ===Materials=== | ||
<u>Stimulation</u> | |||
* hTERT (Airway smooth muscle cells, ASMC's) (D12 P27/D9 P32; split [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/12| 12Feb2010]], put to S<sup>0</sup> [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/15|15Feb2010]]) | * hTERT (Airway smooth muscle cells, ASMC's) (D12 P27/D9 P32; split [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/12| 12Feb2010]], put to S<sup>0</sup> [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/15|15Feb2010]]) | ||
* [http://www.promega.com/catalog/catalogproducts.aspx?categoryname=productleaf_1422 InCELLect™ St-<b>Ht31P</b> Control Peptide] | * [http://www.promega.com/catalog/catalogproducts.aspx?categoryname=productleaf_1422 InCELLect™ St-<b>Ht31P</b> Control Peptide] | ||
* Warm [http://products.invitrogen.com/ivgn/product/42430025 DMEM] (S<sup>0</sup>) | * Warm [http://products.invitrogen.com/ivgn/product/42430025 DMEM] (S<sup>0</sup>) | ||
* Warm [[PBS]] | * Warm [[PBS]] | ||
*DMEM S<sup>0</sup> | |||
** 500 mL DMEM | |||
** 3 mL Fungizone | |||
** 10 mL PenStrep | |||
<u>ELISA</u> | |||
* Supernatants collected [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/05|5March2010]] (D12), [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/10|10March2010]] (D12, P25) & [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/11|11March2010]] (D9, P21) | |||
* [[PBS]] 5% | * [[PBS]] 5% | ||
* [http://en.wikipedia.org/wiki/Bovine_serum_albumin BSA] 5% | * [http://en.wikipedia.org/wiki/Bovine_serum_albumin BSA] 5% in PBS (20 mL per 96-wells) | ||
* [http://www.sanquin.pl/datasheet/m1918.pdf Pelikine Compact™ Human IL-8 ELISA kit] | * [http://www.sanquin.pl/datasheet/m1918.pdf Pelikine Compact™ Human IL-8 ELISA kit] | ||
* Washing buffer (PBS + 0.005% TWEEN 20) | * Washing buffer (PBS + 0.005% TWEEN 20) | ||
Line 38: | Line 41: | ||
** 30 mg [http://en.wikipedia.org/wiki/3,3%E2%80%99,5,5%E2%80%99-Tetramethylbenzidine TMB] | ** 30 mg [http://en.wikipedia.org/wiki/3,3%E2%80%99,5,5%E2%80%99-Tetramethylbenzidine TMB] | ||
** 5 mL [http://en.wikipedia.org/wiki/Dimethyl_sulfoxide DMSO] | ** 5 mL [http://en.wikipedia.org/wiki/Dimethyl_sulfoxide DMSO] | ||
* | * Dilution buffer (1:5, 32 mL H<sub>2</sub>O, 8 mL undiluted buffer) | ||
===Method=== | ===Method=== | ||
Line 47: | Line 47: | ||
* Remove medium form 24-wells plate with ASMC's grown ON in DMEM (S<sup>0</sup>) | * Remove medium form 24-wells plate with ASMC's grown ON in DMEM (S<sup>0</sup>) | ||
* Rinse twice with warm PBS and remove buffer | * Rinse twice with warm PBS and remove buffer | ||
* Add | * Add 300 μL DMEM S<sup>0</sup> with HT31P 0 - 50 μM (see schedule below) | ||
** Incubate 20 min. | ** Incubate 20 min. | ||
** Prepare 25 mL DMEM S<sup>0</sup> with 100% CSE | ** Prepare 25 mL DMEM S<sup>0</sup> with 100% CSE | ||
** Dilute 100% CSE to | ** Dilute 100% CSE to 30% CSE (3 mL CSE 100% + 7 mL S<sup>0</sup>) | ||
* Add | * Add 300 μL 30% CSE to wells according to schedule below (lane C & D) | ||
* Incubate 24 h. | * Incubate 24 h. | ||
Line 61: | Line 57: | ||
|+'''Stimulation set-up''' | |+'''Stimulation set-up''' | ||
! | ! | ||
!{{todo|HT31P}}<BR><u>1</u> | !{{todo|HT31P}}<BR>0 μM<BR><u>1</u> | ||
!{{todo|HT31P}}<BR><u>2</u> | !{{todo|HT31P}}<BR>10 μM<BR><u>2</u> | ||
!{{todo|HT31P}}<BR><u>3</u> | !{{todo|HT31P}}<BR>20 μM<BR><u>3</u> | ||
!{{ | !{{todo|HT31P}}<BR>30 μM<BR><u>4</u> | ||
!{{ | !{{todo|HT31P}}<BR>40 μM<BR><u>5</u> | ||
!{{ | !{{todo|HT31P}}<BR>50 μM<BR><u>6</u> | ||
|- | |- | ||
|<b>A</b><BR> (CTR) | |<b>A</b><BR> (CTR) | ||
|style="background: | |style="background:#FF0000"| | ||
|style="background: | |style="background:#F62217"| | ||
|style="background: | |style="background:#E41B17"| | ||
|style="background: | |style="background:#F62817"| | ||
|style="background: | |style="background:#E42217"| | ||
|style="background: | |style="background:#C11B17"| | ||
|- | |- | ||
|<b>B</b><BR> ( | |<b>B</b><BR> (CTR) | ||
|style="background: | |style="background:#FF0000"| | ||
|style="background: | |style="background:#F62217"| | ||
|style="background: | |style="background:#E41B17"| | ||
|style="background: | |style="background:#F62817"| | ||
|style="background: | |style="background:#E42217"| | ||
|style="background: | |style="background:#C11B17"| | ||
|- | |- | ||
|<b>C</b><BR> (CSE | |<b>C</b><BR> (CSE) | ||
|style="background: | |style="background:#87F717"| | ||
|style="background: | |style="background:#5FFB17"| | ||
|style="background: | |style="background:#59E817"| | ||
|style="background: | |style="background:#7FE817"| | ||
|style="background: | |style="background:#4CC417"| | ||
|style="background: | |style="background:#6CC417"| | ||
|- | |- | ||
|<b>D</b><BR> (CSE | |<b>D</b><BR> (CSE) | ||
|style="background: | |style="background:#87F717"| | ||
|style="background: | |style="background:#5FFB17"| | ||
|style="background: | |style="background:#59E817"| | ||
|style="background: | |style="background:#7FE817"| | ||
|style="background: | |style="background:#4CC417"| | ||
|style="background: | |style="background:#6CC417"| | ||
|- | |- | ||
|} | |} | ||
<b>CTR = control, CSE = Cigarette Smoke extract | <b>CTR = control, CSE = Cigarette Smoke extract</b> | ||
====[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/18|ELISA]]==== | ====[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/18|ELISA]]==== | ||
<b><u>Preparing Sandwich</u></b> | <b><u>Preparing Sandwich</u></b> | ||
Line 176: | Line 173: | ||
* Stop reaction, add 100 μL stopping solution | * Stop reaction, add 100 μL stopping solution | ||
** It will turn yellow | ** It will turn yellow | ||
* Measure absorbance @ | * Measure absorbance @ 450 nm | ||
** Check duplo standard curve | ** Check duplo standard curve | ||
** Check if values fall within standard curve (preferred) | ** Check if values fall within standard curve (preferred) | ||
Line 305: | Line 302: | ||
| | | | ||
| | | | ||
| | |BLANC | ||
| | |BLANC | ||
|- | |- | ||
|} | |} | ||
====Co-immunoprecipitation==== | ====Co-immunoprecipitation==== | ||
* resuspend beads prepared [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/15| 15March2010]] | * resuspend beads prepared [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/15| 15March2010]] | ||
Line 319: | Line 317: | ||
* Incubate ON @ °C | * Incubate ON @ °C | ||
====Splitting cells==== | |||
* D9 P23 & D12 P27 were split, one petridish to 4 (P+1) | |||
==Results== | ==Results== | ||
{| border="1" | |||
|+'''96-wells plate ELISA''' | |||
|<center>#</center> | |||
!01 <br><b>S<sup>0</sup></b> | |||
!02 <br><b>S<sup>0</sup></b> | |||
!03 <br><b>S<sup>0</sup></b> | |||
!04 <br><b>Ht31</b> | |||
!05 <br><b>Ht31</b> | |||
!06 <br><b>Ht31</b> | |||
!07 <br><b>S<sup>0</sup></b> | |||
!08 <br><b>S<sup>0</sup></b> | |||
!09 <br><b>S<sup>0</sup></b> | |||
!10 <br><b>Ht31</b> | |||
!11 <br><b>Ht31</b> | |||
!12 <br><b>Ht31</b> | |||
|- | |||
!A <br><b>CTR</b> | |||
|style="background:red; color:white" | 0,328 | |||
|style="background:red; color:white" |0,555 | |||
|style="background:red; color:white" |0,680 | |||
|style="background:red; color:white" |1,051 | |||
|style="background:red; color:white" |1,127 | |||
|style="background:red; color:white" |1,024 | |||
|style="background:blue; color:white" |0,187 | |||
|style="background:blue; color:white" |0,159 | |||
|style="background:blue; color:white" |0,133 | |||
|style="background:blue; color:white" |0,156 | |||
|style="background:blue; color:white" |0,182 | |||
|style="background:blue; color:white" |0,182 | |||
|- | |||
!B <br><b>CSE</b> | |||
|style="background:red; color:white" |0,836 | |||
|style="background:red; color:white" |1,341 | |||
|style="background:red; color:white" |1,476 | |||
|style="background:red; color:white" |1,188 | |||
|style="background:red; color:white" |1,093 | |||
|style="background:red; color:white" |1,261 | |||
|style="background:blue; color:white" |1,623 | |||
|style="background:blue; color:white" |1,367 | |||
|style="background:blue; color:white" |1,436 | |||
|style="background:blue; color:white" |0,819 | |||
|style="background:blue; color:white" |1,111 | |||
|style="background:blue; color:white" |0,856 | |||
|- | |||
!C <br><b>CSE+8P</b> | |||
|style="background:red; color:white" |1,148 | |||
|style="background:red; color:white" |1,229 | |||
|style="background:red; color:white" |1,227 | |||
|style="background:red; color:white" |0,999 | |||
|style="background:red; color:white" |1,185 | |||
|style="background:red; color:white" |1,172 | |||
|style="background:blue; color:red" |2,237 | |||
|style="background:blue; color:white" |1,332 | |||
|style="background:blue; color:white" |1,613 | |||
|style="background:blue; color:white" |1,456 | |||
|style="background:blue; color:white" |1,158 | |||
|style="background:blue; color:white" |1,379 | |||
|- | |||
!D <br>CSE+6Bnz | |||
|style="background:red; color:white" |0,530 | |||
|style="background:red; color:white" |0,546 | |||
|style="background:red; color:white" |0,496 | |||
|style="background:red; color:white" |0,388 | |||
|style="background:red; color:white" |0,295 | |||
|style="background:red; color:white" |0,396 | |||
|style="background:blue; color:white" |0,458 | |||
|style="background:blue; color:white" |0,394 | |||
|style="background:blue; color:white" |0,368 | |||
|style="background:blue; color:white" |0,288 | |||
|style="background:blue; color:white" |0,284 | |||
|style="background:blue; color:white" |0,290 | |||
|- | |||
!E <br>CTR | |||
|style="background:orange; color:white" |0,175 | |||
|style="background:orange; color:white" |0,185 | |||
|style="background:orange; color:white" |0,160 | |||
|style="background:orange; color:white" |0,213 | |||
|style="background:orange; color:white" |0,268 | |||
|style="background:orange; color:white" |0,236 | |||
|1,958 | |||
|1,374 | |||
|0,709 | |||
|0,355 | |||
|0,180 | |||
|0,119 | |||
|- | |||
!F <br>CSE | |||
|style="background:orange; color:white" |0,280 | |||
|style="background:orange; color:white" |0,704 | |||
|style="background:orange; color:white" |0,525 | |||
|style="background:orange; color:white" |0,553 | |||
|style="background:orange; color:white" |0,571 | |||
|style="background:orange; color:white" |0,650 | |||
|1,815 | |||
|1,359 | |||
|0,705 | |||
|0,404 | |||
|0,192 | |||
|0,129 | |||
|- | |||
!G <br>CSE+8P | |||
|style="background:orange; color:white" |0,464 | |||
|style="background:orange; color:white" |0,516 | |||
|style="background:orange; color:white" |0,526 | |||
|style="background:orange; color:white" |0,427 | |||
|style="background:orange; color:white" |0,589 | |||
|style="background:orange; color:white" |0,741 | |||
|0,089 | |||
|0,064 | |||
|0,071 | |||
|0,073 | |||
|0,070 | |||
|0,086 | |||
|- | |||
!H <br>CSE+6Bnz | |||
|style="background:orange; color:white" | 0,378 | |||
|style="background:orange; color:white" |0,220 | |||
|style="background:orange; color:white" |0,214 | |||
|style="background:orange; color:white" |1,733 | |||
|style="background:orange; color:white" |0,326 | |||
|style="background:orange; color:white" |0,222 | |||
|0,091 | |||
|0,071 | |||
|0,071 | |||
|0,013 | |||
|0,014 | |||
|0,034 | |||
|- | |||
|} | |||
<BR> | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"|pg/ml | |||
| align="center" style="background:#f0f0f0;"|pg/ml | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"|pg/mL | |||
|- | |||
| align="center" style="background:#f0f0f0;"|'''D12''' | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"|'''aver OD''' | |||
| align="center" style="background:#f0f0f0;"|'''Aver Il-8''' | |||
| align="center" style="background:#f0f0f0;"|'''Dilu''' | |||
| align="center" style="background:#f0f0f0;"|'''alamar blue''' | |||
| align="center" style="background:#f0f0f0;"|'''% to control''' | |||
| align="center" style="background:#f0f0f0;"|'''Corrected''' | |||
|- | |||
| S0||CTR||0,618||32,1||64,1||4088,3||100,0||64,1 | |||
|- | |||
| ||CSE||1,409||79,5||318,1||5517,0||134,9||235,7 | |||
|- | |||
| ||CSE+8-p||1,201||67,1||268,4||5710,3||139,7||192,2 | |||
|- | |||
| ||CSE+Bnz||0,524||26,5||105,9||4982,7||121,9||86,9 | |||
|- | |||
| HT31||CTR||1,067||59,1||118,1||6815,0||166,7||70,9 | |||
|- | |||
| ||CSE||1,181||65,9||526,9||4659,0||114,0||462,3 | |||
|- | |||
| ||CSE+8-p||1,119||62,1||497,1||4882,0||119,4||416,3 | |||
|- | |||
| ||CSE+Bnz||0,360||16,6||132,8||4353,0||106,5||124,8 | |||
|} | |||
<BR> | |||
[[Image:IL8secretionD12PLATE116032010.JPG.JPG]] | |||
<BR> | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"|pg/ml | |||
| align="center" style="background:#f0f0f0;"|pg/ml | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"|pg/mL | |||
|- | |||
| align="center" style="background:#f0f0f0;"|'''D12 P25''' | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"|'''aver OD''' | |||
| align="center" style="background:#f0f0f0;"|'''Aver Il-8''' | |||
| align="center" style="background:#f0f0f0;"|'''Dilu''' | |||
| align="center" style="background:#f0f0f0;"|'''alamar blue''' | |||
| align="center" style="background:#f0f0f0;"|'''% to control''' | |||
| align="center" style="background:#f0f0f0;"|'''Corrected''' | |||
|- | |||
| S0||CTR||0,160||4,6||9,2||8507,3||100,0||9,2 | |||
|- | |||
| ||CSE||1,475||83,5||334,1||6568,7||77,2||432,8 | |||
|- | |||
| ||CSE+8-p||1,473||83,4||333,5||6875,7||80,8||412,6 | |||
|- | |||
| ||CSE+Bnz||0,407||19,4||77,7||5567,3||65,4||118,7 | |||
|- | |||
| HT31||CTR||0,173||5,4||10,9||8273,7||97,3||11,2 | |||
|- | |||
| ||CSE||0,838||45,3||362,2||5215,3||61,3||590,8 | |||
|- | |||
| ||CSE+8-p||1,331||74,9||599,0||5988,7||70,4||850,9 | |||
|- | |||
| ||CSE+Bnz||0,287||12,3||98,1||2887,0||33,9||289,2 | |||
|} | |||
<BR> | |||
[[Image:IL8secretionD12PLATE216032010.JPG]] | |||
<BR> | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"|pg/ml | |||
| align="center" style="background:#f0f0f0;"|pg/ml | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"|pg/mL | |||
|- | |||
| align="center" style="background:#f0f0f0;"|''' D9 P21''' | |||
| align="center" style="background:#f0f0f0;"| | |||
| align="center" style="background:#f0f0f0;"|'''aver OD''' | |||
| align="center" style="background:#f0f0f0;"|'''Aver Il-8''' | |||
| align="center" style="background:#f0f0f0;"|'''Dilu''' | |||
| align="center" style="background:#f0f0f0;"|'''alamar blue''' | |||
| align="center" style="background:#f0f0f0;"|'''% to control''' | |||
| align="center" style="background:#f0f0f0;"|'''Corrected''' | |||
|- | |||
| S0||CTR||0,173||5,4||10,9||10141,3||100,0||10,9 | |||
|- | |||
| ||CSE||0,503||25,2||100,8||9616,0||94,8||106,3 | |||
|- | |||
| ||CSE+8-p||0,502||25,1||100,6||10673,0||105,2||95,6 | |||
|- | |||
| ||CSE+Bnz||0,271||11,3||45,1||11619,7||114,6||39,3 | |||
|- | |||
| HT31||CTR||0,239||9,4||18,7||8867,3||87,4||21,4 | |||
|- | |||
| ||CSE||0,591||30,5||244,0||9047,0||89,2||273,5 | |||
|- | |||
| ||CSE+8-p||0,586||30,2||241,3||9436,7||93,1||259,3 | |||
|- | |||
| ||CSE+Bnz||0,274||11,5||91,7||11161,3||110,1||83,3 | |||
|- | |||
| | |||
|} | |||
<BR> | |||
[[Image:IL8secretionD9PLATE316032010.JPG]] | |||
==Conclusion== | ==Conclusion== | ||
*{{done}} Ow.. | *{{done}} Ow.. | ||
==Related== | ==Related entries== | ||
* [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/12| Splitting cells]] | ====Run 4: Ht31P dose/response curve D9/D12==== | ||
* [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/15| Cells put to S<sup>0</sup>]] | * [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/12| Splitting cells 12March2010]] | ||
* [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/15| Cells put to S<sup>0</sup> 15March2010]] | |||
====Run 3: Ht31/S<sup>0</sup>==== | |||
*Counting cells | |||
**[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/26|26Feb2010 D9/D12]] | |||
**[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/05|03March2010 D9 (P21)/D12 (P25)]] | |||
*Putting to S<sup>0</sup> | |||
**[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/03|03March D9/D12]] | |||
**[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/08|08March2010 D12 (P25)]] | |||
**[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/09|09March2010 D9 (P21)]] | |||
*Stimulating cells | |||
**[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/04|04March2010 D9 (samples lost)/D12]] | |||
**[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/09|09March2010 D12 (P25)]] | |||
**[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/10|10March2010 D9 (P21)]] | |||
*Ending stimulation & Viability | |||
**[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/05|05March2010 D12]] | |||
**[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/10|10March2010 D12 (P25)]] | |||
**[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/11|11March2010 D9 (P21)]] | |||
*ELISA | |||
**[[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/03/16|16March2010 D12, D12 (P25) & D9 (P21)]] | |||
==Same actions== | ==Same actions== | ||
* [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/16| Cell stimulation]] | * [[User:Wilfred_J._Poppinga/Notebook/cAMP_compartmentalization/2010/02/16| Cell stimulation]] | ||
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</html> [[Media:HTERT_Donor_12_ht31_160310.xls| Excel hTERT D12]] | |||
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</html> [[Media:HTERT_Donor_9_P21_160310.xls| Excel hTERT D9 P21]] | |||
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Revision as of 14:38, 18 March 2010
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ELISA & hTERT stimulation | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Summary
Materials & MethodsMaterialsStimulation
ELISA
MethodStimulation hTERT cells
CTR = control, CSE = Cigarette Smoke extract ELISAPreparing Sandwich
Co-immunoprecipitation
(possible also beads (30 μL) with only sample (200 μL))
Splitting cells
Results
Conclusion
Related entriesRun 4: Ht31P dose/response curve D9/D12Run 3: Ht31/S0
Same actionsAttachment<html> <a href="http://openwetware.org/wiki/Image:HTERT_Donor_12_ht31_160310.xls"> <img src="http://openwetware.org/images/f/f8/Owwnotebook_icon.png" alt="" width="50" height="50"> </a> </html> Excel hTERT D12 <html> <a href="http://openwetware.org/wiki/Image:HTERT_donor_12_P25_160310.xls"> <img src="http://openwetware.org/images/f/f8/Owwnotebook_icon.png" alt="" width="50" height="50"> </a> </html> Excel hTERT D12 P25 <html> <a href="http://openwetware.org/wiki/Image:HTERT_Donor_9_P21_160310.xls"> <img src="http://openwetware.org/images/f/f8/Owwnotebook_icon.png" alt="" width="50" height="50"> </a> </html> Excel hTERT D9 P21 |