Gel Running Buffer.docx

1、Gel Running BufferGel Running BufferAlthough Tris-glycine SDS-PAGE gels are the most common method for separating proteins for analysis, any gel separation technique can be used. Other techniques for protein separation include: isoelectric focusing, SDS-PAGE using buffers other than Tris-glycine, ur

2、ea-polyacrylamid gels, tris-borate and native low-salt polyacrylamide gels. All the buffers are prepared with high quality reagents. Electrophoresis buffers are sold as concentrate. Please dilute with deionized water to make 1X. Please Note: The composition of 1X is provided unless specified otherwi

3、se.M0195 - MES SDS Running Buffer. 500 ml, sterile solution. 500 mlDescription: 3.2g MES (2-N-Morpholinoethanesulfonic acid) monohydrate, 121.2g Tris Base, 20.0g Sodium Dodecyl Sulfate, 7.631g EDTA.ProductCatalog No.QuantityPriceIEF Anode buffer (50X), pH 2.4Phosphoric acid 7 mMBP-210500 ml1000 ml$1

4、5$25IEF Cathode buffer (10X) pH 10.1Arginine (free base20 mMLysine (free base) 20 mMBP-211250 ml500 ml$35$65IEF Cathode buffer (10X) pH 3-7Lysine (free base) 20 mMBP-214250 ml500 ml$35$65MES-SDS running buffer (20X), pH 7.3 MES50 mMTris base 50 mMSDS0.1%EDTA 1 mM BP-177500 ml1000 ml$42$80MOPS-SDS ru

5、nning buffer (20X), pH 7.7MOPS50 mMTris base50 mMSDS0.1%EDTA1 mM BP-178500 ml1000 ml$42$80Tris-Acetate-SDS running buffer (10X), pH 8.24Tris base 50 mMTricine50 mMSDS3.5 mMBP-179500 ml1000 ml$30$55Tris-Glycine-SDS running buffer (10X), pH 8.3Tris-base 25 mMGlycine192 mMSDS 0.1% BP-1501000 ml2000 ml4

6、000 ml$21$37$70Tris-Glycine-native running buffer (10X), pH 8.5Tris-base25 mMGlycine192 mM BP-1601000 ml2000 ml4000 ml$21$37$70Tris-Tricine-SDS running buffer (10X), pH 8.3, (Cathode buffer)Tris base 100 mMTricine 100 mMSDS 0.1% BP-170500 ml1000 ml$37$70Tris-Tricine-SDS running buffer (10X), pH 8.9,

7、 (Anode buffer)Tris-HCl 200 mMBP-172500 ml1000 ml$21$37Tris-Tricine-SDS buffer (10X) kitThe kit contains one each of BP-170 and BP-172BP-175500 ml1000 ml$45$85ProductCatalog No.QuantityPriceTBE running buffer (10X), pH 8.3Tris base 89 mMBoric acid 89 mMEDTA 2 mM BP-1801000 ml2000 ml4000 ml$21$37$70Z

8、ymogram Renaturation buffer (10X)TritonX100 2.7% (w/v) in DI waterBP-217500 ml1000 ml$15$25Zymogram Developing buffer (10X)Tris-HCl (pH, 7.5) 50 mMNaCl 200 mMCacl25 mMBrij 35 0.02%BP-218500 ml1000 ml$21$37Blot Transfer BufferTransfer of proteins from the gel onto the membrane is achieved in three wa

9、ys: simple diffusion, vacuum-assisted solvent flow, and electrophoretic elution. Electrophoretic elution is the method of choice because of its ease, speed and completeness of transfer. Electrophoretic elution can be achieved either by wet or by semi-dry transfer. Please Note: The composition of 1X

10、is provided unless specified otherwise.ProductCatalog No.QuantityPriceTransfer or Electro blotting buffer (10X), pH 8.3Tris-base 25 mMGlycine192 mMJust add methanol according to your protocolBP-1901000 ml2000 ml4000 ml$21$37$70Tris-Caps buffer (10X)Anode buffer, pH 9.6 Cathode buffer, pH 9.5Tris60 m

11、MTris60 mMCaps 40 mMCaps 40 mMMethanol (add) 15% SDS0.1%BP-192500 ml1000 ml$25$45Luminol and Oxidizing Solutions for western blotting (HRP Based) Just mix Luminol and Oxidizing solution, incubate the membrane for 1 min. and expose the x-ray film for different periods of time. WB-100 250 ml each 500

12、ml each $50 $85 Luminol and Oxidizing Solutions for western blotting (HRP Based) Just mix Luminol and Oxidizing solution, incubate the membrane for 1 min. and expose the x-ray film for different periods of time.WB-100250 ml each500 ml each$50$85Blot Blocking BufferBefore the blot can be processed fo

13、r antigen detection, it is very important to block the non-specific binding site for the antibodies. BBP provides following reagents that are used to block the non-specific sites on the membrane.Please Note: The composition of 1X is provided unless specified otherwise.ProductCatalog No.QuantityPrice

14、BSA-blocking buffer in PBS3% BSA in PBSIBB-100200ml500ml$30$55BSA-blocking buffer in TBS 3% BSA in TBSIBB-110200 ml500 ml$30$55Casein-blocking buffer in PBS3% Casein in PBSIBB-120500 ml1000 ml$30$55Casein-blocking buffer in PBS (with azide)3% Casein and 0.02% Sodium azide in PBSIBB-121500 ml1000 ml$

15、30$55ProductCatalog No.QuantityPriceCasein-blocking buffer in TBS 3% Casein in TBSIBB-130500 ml1000 ml$30$55Casein-blocking buffer in TBS (with azide)3% Casein and 0.02% Sodium azide in TBSIBB-131500 ml1000 ml$30$55Gelatin-blocking buffer in PBS1% Gelatin in PBSIBB-134500 ml1000 ml$30$55Gelatin-bloc

16、king buffer in PBS (with azide)1% Gelatin and 0.02% Sodium azide in PBSIBB-135500 ml1000 ml$30$55Gelatin-blocking buffer in TBS1% Gelatin in TBSIBB-136500 ml1000 ml$30$55Gelatin-blocking buffer in TBS (with azide)1% Gelatin and 0.02% Sodium azide in TBSIBB-137500 ml1000 ml$30$55Blotto in PBS5% nonfa

17、t dry milk, 0.02% Sodium azide in PBSIBB-140250 ml500 ml$25$40Blotto in TBS5% nonfat dry milk, 0.02% Sodium azide in TBSIBB-150250 ml500 ml$25$40Blotto/Tween in TBS5% nonfat dry milk, 0.1% Tween-20, 0.02% Sodium azide in TBS IBB-160500 ml1000 ml$25$40PBS/Tween-20 (10X)0.1% Tween-20 in PBS IBB-170500

18、 ml1000 ml$25$40PBS/Tween-20 (10X)0.05% Tween-20 in PBS IBB-171500 ml1000 ml$25$40TBS/Tween-20 (10X)0.1% Tween-20 in TBS IBB-180500 ml1000 ml$25$40TBS/Tween-20 (10X)0.05% Tween-20 in TBS IBB-181500 ml1000 ml$25$40Gelatin 10%IBB-184100 ml200 ml$35$60Gelatin/Tween in PBS1% Gelatin, 0.05% Tween-20, 0.0

19、2% Sodium azide in PBS IBB-185500 ml1000 ml$40$70Gelatin/Tween in TBS1% Gelatin, 0.05% Tween-20, 0.02% Sodium azide in TBS IBB-186500 ml1000 ml$40$70Horse Serum10% Horse serum, 0.02% sodium azide in PBSIBB-190100 ml200 ml$35$60Blot Washing Buffer Please Note: The composition of 1X is provided unless

20、 specified otherwise.ProductCatalog No.QuantityPricePhosphate-buffered saline (PBS, 10X, pH 7.4) Phosphate buffer 10 mMKCL2.7 mMNaCl 137 mM BM-2201000 ml2000 ml4000 ml$21$37$70PBS-Tween, (10X, pH 7.4) Phosphate buffer 10 mMNaCl 150 mMTween-20 0.05% BM-221250 ml500 ml$21$37Phosphate-buffered saline (

21、PBS, 10X, pH 7.4)For western blot washingSodium Phosphate buffer 10 mMNaCl150 mM BM-2221000 ml2000 ml4000 ml$21$37$70Tris-buffered saline (TBS, 10X)Tris25 mMKCL2.7 mMNaCl137 mMBM-3001000 ml2000 ml4000 ml$21$37$70TBS-Tween-20, (10X. pH 7.4)Tris50 mMNaCl150 mMTween-20 0.05% IBB-181250 ml500 ml$21$37Tr

22、is-buffered saline (TBS, 10X, pH 7.4)For western blot washingTris 50 mMNaCl 150 mM BM-3011000 ml2000 ml4000 ml$21$37$70Tris-buffered saline (10X, high salt, pH 7.4)For western blot washingTris50 mMNaCl500 mMBM-301A1000 ml2000 ml4000 ml$25$45$80Tris-buffered saline (TBS, 10X, pH 8.0)Tris50 mMNaCl 150

23、 mM BM-301B1000 ml2000 ml4000 ml$21$37$70Blot Stripping BufferProductCatalog No.QuantityPriceStripping buffer, (4X), pH 6.8Tris-HCl 250 mMSDS8%Just add mercaptoethnolBP-96250 ml500 ml$25$45Stripping buffer 2, (4X), pH 2.5Glycine0.2 MBP-97250 ml500 ml$25$45MOLECULAR BIOLOGY: WORKING WITH PROTEINS PRO

24、TEIN DETECTION: GEL ELECTROPHORESIS RUNNING NOVEX NuPage GELS Running Novex NuPage Gels Title | Overview | Procedure | Solutions | BioChemicals | Hints | Printable Version Contributor:The Laboratory of Steve Hahn at the Fred Hutchinson Cancer Research Center OverviewTitle | Overview | Procedure | So

25、lutions | BioChemicals | Hints | Printable Version These are simplified instructions for use of Novex precast NuPage gels. The XCEll II mini cell apparatus is required for this protocol. For more information on Novex gels and products see their webpage ProcedureTitle | Overview | Procedure | Solutions | BioChemicals | Hints | Printable Ve