Overview: Difference between revisions

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=== Antibody ===
=== Antibody ===
===== Monoclonal Antibodies: =====
===== List of primary antibodies =====
{| class="wikitable"
| 12/101 ||  || Developmental Studies Hybridoma Bank ||  || Mouse ||  ||  || Yuka || 4°C || 46 ug/ml Ig || 1:200 || 1:200 ||  ||  ||  ||
|-
| 15.3B9 (NOT1) ||  || Developmental Studies Hybridoma Bank || 15.3B9 (NOT1) || Mouse || Monoclonal || notochord marker (Chicken) || Yuka || 4°C ||  || 21 ug/ml IG ||  ||  ||  || http://dshb.biology.uiowa.edu/notochord-marker ||
|-
| 20B4 ||  || Developmental Studies Hybridoma Bank || 20B4 || Mouse || Monoclonal || neural crest cell (Chicken) || Yuka || 4°C ||  ||  ||  ||  ||  || http://dshb.biology.uiowa.edu/neural-crest-cells ||
|-
| 5D3 ||  || Developmental Studies Hybridoma Bank || 5D3 || Mouse || Monoclonal || Cadherin E (Xenopus) || Yuka || 4°C ||  || 37 ug/ml IG ||  ||  ||  || http://dshb.biology.uiowa.edu/5D3 ||
|-
| 8C8 ||  || Developmental Studies Hybridoma Bank || 8C8 || Mouse || Monoclonal || integrin beta-1/ CD29 (Xenopus) || Yuka || 4°C ||  ||  ||  ||  ||  || http://dshb.biology.uiowa.edu/integrin-beta-1_7 ||
|-
| Aggrecan ||  || millipore || AB1031 || Rabbit ||  ||  || Lidia ||  ||  ||  || Not work, Dim || '1:500 (Lidia) ||  ||  ||
|-
| Alpha tubulin (DM1a) ||  || MPI ||  || Mouse || Monoclonal || Chicken alpha tubulin || Takuji || -20°C ||  ||  ||  ||  || 1:1000 -  ||  ||
|-
| b-catenin ||  ||  Gift from Thomas Kurth || P14L || Rabbit ||  || Xenopus || Yuka_Aliquot from Tomas Kurth ||  ||  ||  ||  ||  ||  || Beta-catenin translocation into nuclei demarcates the dorsalizing centers in frog and fish embryos. ||
|-
| Bra ||  ||  || AF2085 ||  ||  ||  || Yuka || -20C ||  || 0.2ug/ml in PBS ||  ||  ||  ||  ||
|-
| BrdU (Bu20a) ||  || MPI ||  || Mouse || Monoclonal || BrdU - thymidine analog || Takuji || -20°C ||  || 1:400 -  ||  ||  ||  ||  ||
|-
| BrdU ||  || Antibodies-online || ABIN964591 || Rabbit || Polyclonal || BrdU - thymidine analog || Takuji || -20°C ||  || 1:600 - ||  ||  ||  || http://www.antibodies-online.com/antibody/964591/anti-Bromodeoxyuridine+BrDU+antibody/ ||
|-
| Casp3 ||  || Abcam || ab13847 || rabbit ||  ||  || Lidia || -20C ||  ||  ||  ||  ||  ||  ||
|-
| col1A2 ||  || Developmental Studies Hybridoma Bank || SP1.D8 || Mouse || Monoclonal || Sheep || Yuka || -20C || 187ug/ml || 1:200- || 1:200_found in most connective tissue and abundant in bone, cornea, dermis and tendon. ||  ||  ||  ||
|-
| col2A1 ||  || Acris || AM00619PU-N || Mouse ||  ||  || Yuka || -20C || 0.2mg/ml in /H2O || 1:10 || 1:100-200 ||  ||  ||  ||
|-
| Digoxigenin-AP ||  || Roche || 11093274910 || Sheep || Polyclonal ||  || Common box || 4°C ||  ||  ||  ||  ||  || http://www.sigmaaldrich.com/catalog/product/roche/11093274910?lang=de&region=AT&gclid=Cj0KEQjwk-jGBRCbxoPLld_bp-IBEiQAgJaftdEIkv2iCvPxqx7NfQns9MUkipGFBChRXSL1FhsYOxkaApPb8P8HAQ ||
|-
| FITC ||  || Jackson ImmunoResearch || 200-002-037 || Mouse || Monoclonal || Fluorescein isothiocyanate || Takuji || -20°C ||  || 1:400 -  ||  ||  ||  || https://www.jacksonimmuno.com/catalog/products/200-002-037 ||
|-
| FITC ||  || invitrogen || 71-1900 || Rabbit || Polyclonal || Fluorescein isothiocyanate || Takuji || -20°C ||  || 1:400 -  ||  ||  ||  || https://www.thermofisher.com/antibody/product/FITC-Antibody-Polyclonal/71-1900 ||
|-
| GFP ||  || Rockland || 600-401-215 || Rabbit ||  ||  || Common box || -20C ||  ||  || 1:2000-4000 ||  ||  ||  ||
|-
| GFP ||  || MPI ||  || Goat ||  ||  || Yuka (from Jifeng) || -20C ||  || Yuka (from Jifeng) || 1:2000- ||  ||  ||  ||
|-
| GFP ||  || MPI || 106-A20-Mix || Mouse ||  ||  || Yuka || -20C || 2.96 mg/ml in PBS ||  || 1:200, High background ||  ||  ||  ||
|-
| GFP ||  || Abcam || ab13970 || Chick ||  ||  || Yuka (from Andrea), Lidia || -20C ||  || Andrea || High background ||  ||  ||  ||
|-
| GFP ||  || Torrey Pines || TP401 || Rabbit ||  ||  || Yuka || -20C ||  || 1:200 (Axolotl) ||  ||  ||  ||  ||
|-
| GFP ||  || Invitrogen || A11120 || Mouse || monoclonal ||  || Common box || -20C || 1 mg/ml in PBS ||  ||  ||  ||  ||  ||
|-
| Hoxa11 ||  || Tanaka Lab (Kathleen) ||  || Rabbit || Polyclonal  || Axolotl || Yuka_Aliquot from Kathleen || -20C || 0.95 mg/ml ||  || FF+2%MEMFA.Dent's ||  ||  ||  ||
|-
| Hoxa11 ||  || Tanaka Lab (Kathleen) ||  || Mouse || Monoclonal || Axolotl || Yuka_Aliquot from Kathleen || -20C || 4.22 mg/ml ||  ||  ||  ||  ||  ||
|-
| Hoxa13 ||  || Tanaka Lab (Kathleen) ||  || Rabbit || Polyclonal  || Axolotl || Yuka_Aliquot from Kathleen || -20C || 0.15 mg/ml || 01:50 || 1:50, FreshFrozen+Unfixed ||  ||  ||  ||
|-
| Hoxa9 ||  || Tanaka Lab (Kathleen) ||  || Rabbit || Polyclonal  || Axolotl || Yuka_Aliquot from Kathleen || -20C || 1.35 mg/ml ||  || FF+Methanol+ph9AR ||  ||  ||  ||
|-
| Laminin ||  || Sigma || L9393 || Rabbit ||  ||  || Yuka (from Jifeng) || -20C ||  ||  || 1:200, 500_1:25 (paraffin), 1:100 (Slack) ||  ||  ||  ||
|-
| MBP ||  || Genetex || GTX76114 || Rat || Monoclonal || Myelin basic protein (Bovine) || Common box || -20C ||  || 1:200 || 1:200 ||  ||  ||  ||
|-
| Mef2c ||  || Sigma || HPA005533 || Rabbit || Polyclonal  || Human MEF2C (Myocyte enhancer factor 2C) || Aliquot from Takuji ||  ||  || 1:50 (Prayag), 1:300 (Takuji) || 1:5000,10000 (High background on all type of tissue) ||  ||  ||  ||
|-
| Meis 1/2/3 ||  || Millipore || 05-779 || Mouse IgG1 || Monoclonal || mouse Meis 1 (aa 60-390) || Lidia || -20C (30% glycerol) ||  || 1:200 (Takuji?- frozen tissue secitons) ||  || 1:200 (Lidia?) ||  || http://www.merckmillipore.com/AT/de/product/Anti-Meis-1%2F2%2F3-Antibody%2C-clone-9.2.7,MM_NF-05-779?ReferrerURL=https%3A%2F%2Fwww.google.at%2F&bd=1 ||
|-
| MHC ||  || Developmental Studies Hybridoma Bank || A4.1025 || Mouse ||  ||  || Yuka || 4°C ||  || 1:200 || 1:200 ||  ||  ||  ||
|-
| Osterix ||  || abcam || ab22552 || rabbbit ||  ||  || Lidia ||  ||  ||  ||  ||  ||  ||  ||
|-
| Pax7 ||  || MPI ||  || Mouse ||  ||  || Yuka || -20C ||  || 1:100, 200, 400  ||  ||  ||  ||  ||
|-
| PCNA ||  || Santa Cruz || sc-56 AF64 || Alexa Fluor -647  ||  ||  || Lidia ||  ||  ||  || 1:500 ||  ||  ||  ||
|-
| PDGFRa (16A1) - Biotin ||  || Invitrogen || A15732 || Mouse || Monoclonal || Human CD104a || Yuka (from Josh) || 4°C || 1 mg/ml ||  ||  ||  ||  || https://www.thermofisher.com/antibody/product/PDGFRA-Antibody-clone-16A1-Monoclonal/A15732 ||
|-
| Perilipin ||  || R and D || P1873 || rabbit ||  ||  || Lidia ||  ||  ||  ||  ||  ||  ||  ||
|-
| PHH3 ||  || millipore || 06-570 || rabbit || Polyclonal || Linear peptide corresponding to human Histone H3 at Ser10 || Takuji || 4°C || 1 mg/ml || 1:400 -  ||  ||  ||  || https://www.merckmillipore.com/DE/de/product/Anti-phospho-Histone-H3-%28Ser10%29-Antibody%2C-Mitosis-Marker,MM_NF-06-570?ReferrerURL=https%3A%2F%2Fwww.emdmillipore.com%2FCA%2Fen%2Fproduct%2FAnti-phospho-Histone-H3-%2528Ser10%2529-Antibody%252C-Mitosis-Marker%2CMM_NF-06-570 ||
|-
| PPARgamma ||  || Cell signalling || 81B8 || rabbit ||  ||  || Lidia ||  ||  ||  ||  ||  ||  ||  ||
|-
| Prrx1 ||  || Tanaka Lab (Prayag) ||  || Rabbit || Polyclonal || Axolotl N-terminus (1-101 aa) || Lidia, Yuka_Aliquot from Prayag || -20C ||  || 1:200 || 1:200 ||  ||  ||  ||
|-
| RFP ||  || Rockland || 600-401-379 || Rabbit || Polyclonal ||  || Common box || -20C ||  || 1:300-1:1000 for Axolotl || 1:1000 or 1:2000 ||  ||  || http://www.rockland-inc.com/Product.aspx?id=34801 ||
|-
| Rhodamine ||  || Molecular Probes || A-6397 || Rabbit || Polyclonal || Tetramethylrhodamine || Takuji || -20°C ||  || 1:400 -  ||  ||  ||  ||  ||
|-
| Scleraxis ||  || Santa Cruz || D-14, sc-87425 || Goat || Polyclonal Goat IgG ||  || Yuka || 4°C ||  ||  ||  ||  ||  ||  ||
|-
| Shh (H-160) ||  || Santa Cruz || SC-9024 || Rabbit || Polyclonal ||  || Yuka || 4°C ||  || 200 ug/ml ||  ||  ||  ||  ||
|-
| Sox9 ||  || Chemicon || Ab5535 || Rabbit || Polyclonal || Synthetic peptide from human Sox9, aa486-509 (extreme C-Terminus) || Yuka_Aliquot || -20C ||  || 1:500-1:1000 || 1:500 ||  ||  || http://www.merckmillipore.com/DE/en/product/Anti-Sox9-Antibody,MM_NF-AB5535 ||
|-
| Sox9 ||  || R&D || AF3075 || Goat || Polyclonal || E. coli-derived recombinant human SOX9 (Met1-Lys151) || Lidia, Yuka || -20C ||  || 1:100-1:200 || 1:100 ||  ||  || https://www.rndsystems.com/products/human-sox9-antibody_af3075 ||
|-
| Zo-1 ||  || Invitrogen || 33-9100 || Mouse ||  ||  || Yuka (from Akira) || -20C ||  ||  ||  ||  ||  ||  ||
|-
| GADD45B ||  || SIGMA || SAB2108614-100 || Rabbit || Polyclonal || Peptide with sequence: QIHFTLIQSFCCDNDINIVRVSGMQRLAQLLGEPAETQGTTEARDLHCLL || Marco || -20C || 0.5-1 mg/ml ||  ||  ||  ||  ||  ||
|-
| GADD45G ||  || SIGMA || HPA023606 || Rabbit || Polyclonal || MTLEEVRGQDTVPESTARMQGAGKALHELLLSAQRQGCLTAGVYESAKVLNVDPDNVTFCVLAAGEEDEGDIALQIHFTLIQAFCCENDIDIVRVGDVQRLAAIVG || Marco || -20C || 0.05mg/ml ||  ||  ||  ||  ||  ||
|-
| TDG ||  || SIGMA || HPA052263 || Rabbit || Polyclonal || "WKCLFMSGLSEVQLNHMDDHTLPGKYGIGFTNMVERTTPGSKDLSSKEFREGGRILVQKLQKYQPRIAVFNGKCIYEIFSKEVFGVKVKNLEFG
|-
| " || Marco || -20C || 0.1mg/ml ||  ||  ||  ||  ||  ||
|-
| Tet3 ||  || Diagenode || C15410311 || Rabbit || Polyclonal ||  || Marco || -20C || 1 ug/uL ||  ||  ||  ||  ||  ||
|-
| Tet2 ||  || Diagenode || C15410255 || Rabbit || Polyclonal ||  || Marco || -20C || 1 ug/uL ||  ||  ||  ||  ||  ||
|-
| 5-mC ||  || Diagenode || C15200081-100 || Mouse || Monoclonal ||  || Marco || -20C || 1.36 ug/ uL ||  ||  ||  ||  ||  ||
|-
| 5-hmC ||  || Diagenode || C15410205-20 || Rabbit || Polyclonal ||  || Marco || -20C || 3.5 ug/uL ||  ||  ||  ||  ||  ||
|-
| IgG (Rabbit) ||  || Diagenode || C15410206 || Rabbit || Polyclonal ||  || Marco || -20C || 1 ug/uL ||  ||  ||  ||  ||  ||
|-
| p44/42 MAPK (Erk1/2) ||  || Cell signalling || 4696S || Mouse || Monoclonal ||  || Katharina || -20C ||  || 1 : 500 ||  ||  ||  ||  ||
|-
| Phospho-p44/42 MAPK (Erk1/2) ||  || Cell signalling || 4370S || Rabbit || Monoclonal ||  || Katharina || -20C ||  || 1 : 500 ||  ||  ||  ||  ||
|-
| GFAP-Cy3 ||  || Sigma || C9205 || mouse || monoclonal ||  || Katharina || 4°C ||  || 1 : 500 ||  ||  ||  ||  ||
|-
| SV2 ||  || Developmental Studies Hybridoma Bank ||  || mouse || monoclonal ||  || Katharina || 4°C ||  || 1:200- 1:500 ||  ||  ||  ||  ||
|-
| Ctip2 ||  || Abcam || ab18465 || rat || monolonal ||  || Katharina || '-20°C ||  ||  ||  ||  ||  ||  ||
|-
| SATB2 C-terminal ||  || Abcam || ab51502 || mouse || monoclonal ||  || Katharina || -20°C ||  ||  ||  ||  ||  ||  ||
|-
| Glutamine Synthetase, clone GS-6 ||  || Millipore || MAB302 || mouse  || monolonal ||  || Katharina || '-20°C ||  ||  ||  ||  ||  ||  ||
|-
| Parvalbumin ||  || Millipore || MAB1572 || mouse || monolonal ||  || Katharina || '-20°C ||  ||  ||  ||  ||  ||  ||
|-
| Calretinin ||  || Swant || 7697 || rabbit ||  ||  || Katharina || -80°C ||  ||  ||  ||  ||  ||  ||
|-
| Calbindin ||  || Swant || CB38 || rabbit ||  ||  || Katharina || -80°C
|}
 
===== Monoclonal Antibodies =====
<li>[[Purifying IgG1 from Hybridoma Supernatant]]</li>
<li>[[Purifying IgG1 from Hybridoma Supernatant]]</li>
<li>[[Labelling myosin antibody]]</li>
<li>[[Labelling myosin antibody]]</li>
Line 123: Line 258:
<li>[[Anti-BrDU Antibody labeling]]</li>
<li>[[Anti-BrDU Antibody labeling]]</li>


===== Polyclonal Antibodies: =====
===== Polyclonal Antibodies =====
<li>[[Address for making polyclonal antibodies]]</li>
<li>[[Address for making polyclonal antibodies]]</li>
<li>[[Letter to Froppier (French)]]</li>
<li>[[Letter to Froppier (French)]]</li>
Line 131: Line 266:
<li>[[Antibody purification Sox2]]</li>
<li>[[Antibody purification Sox2]]</li>


===== ELISA: =====
===== ELISA =====
<li>[[ELISA]]</li>
<li>[[ELISA]]</li>


Revision as of 16:15, 23 March 2019

Introduction

On the pages listed below you can find most common protocols used in the lab as well as a list of lab duties along with the names of the responsible persons. Please, contact the responsible persons if you have any question, suggestions or troubles.

Axolotl

Maintainance

In this section you will find some general information on how to maintain the animals.

Mating

In this section you will find the information on how to mate axolotls.

Feeding

This section will give you an overview of how to feed the axolotls.

Axolotl in Vitro Fertilization

This section will give you an overview of how to In vitro fertilize the axolotls.

Interesting observations

In the following section you can find some interesting observations concerning the axolotl, its behavior, caveats of the transgenics and many more.

Genotyping (galaxy)

This section section will give you an overview of how to genotype the axolotls.

Lab duties


Protocols

Retrieved from "https://wiki.tanakalab.org/index.php?title=Overview&oldid=909"