T-Pro Biotechnology

Products

T-Pro RIPA Lysis Buffer is ready-to-use as a working 1X solution and requires no further dilution. T-Pro RIPA Lysis Buffer is intended for the extraction of cellular proteins for the efficient lysis of cells and solubilization of protein, while minimizing protein degradation and maintaining protein immunoreactivity and biological activity. We recommend using 1.0 mL of RIPA Lysis Buffer to lyse 0.5 to 5 x 10⁷ adherent mammalian cells. This buffer contains ionic detergents and may not be suitable for kinase enzymes, if these enzymes are easily denatured. Do not add phosphatase inhibitors when preparing lysates for phosphatase assays. 1X T-Pro RIPA lysis buffer consists of 50 mM Tris HCl, 150 mM NaCl, 1.0% (v/v) NP-40, 1% (w/v) Sodium Deoxycholate and 0.1% (w/v) SDS at a pH of 7.4.

This buffer was meticulously prepared using ultrapure reagents dissolved in highly polished pharmaceutical grade deionized water. Protease and phosphatase inhibitors are recommended but not included in product composition.

T-Pro RIPA Lysis Buffer (1X)

50 mM Tris HCl, 150 mM NaCl, 1.0% (v/v) NP-40, 1% (w/v) Sodium Deoxycholate and 0.1% (w/v) SDS at a pH of 7.4

T-Pro RIPA Lysis Buffer (5X)

250 mM Tris HCl, 750 mM NaCl, 5.0% (v/v) NP-40, 5% (w/v) Sodium Deoxycholate and 0.5% (w/v) SDS at a pH of 7.4

Recommended final concentrations of protease inhibitors:

l  1.0mM Phenylmethylsulfonyl fluoride (PMSF),

l  10 µM Leupeptin,

l  10µM Aprotinin,

l  1.0µM Pepstatin. 

l  Recommended final concentrations of phosphatase inhibitors:

l  1.0mM Na3VO4, 1.0 mM NaF.

Gel Casting Buffer

T-Pro Separating or Resolving Buffer (4X)

No. JB03-C001 (500ML/bot)

1.5 M Tris HCl, 0.4% SDS at a pH of 8.8

T-Pro Stacking Buffer (4X)

No. JB03-C002 (500ML/bot)

0.5 M Tris HCl, 0.4% SDS at a pH of 6.8

Sample Buffer

T-Pro Laemmli (SDS sample) Reagent (reducing 4X)

No. JB06-F002 (10ML/bot)

250 mM Tris HCl, 40% Glycerol, 8% Betamercaptoethanol, 0.02% Bromophenol blue and 8% SDS at a pH of 6.8

T-Pro Laemmli (SDS sample) Reagent (non-reducing 4X)

No. JB06-F003 (10ML/bot)

250 mM Tris HCl, 40% Glycerol, 0.02% Bromophenol blue and 8% SDS at a pH of 6.8

T-Pro Laemmli (SDS sample) Reagent (reducing 6X)

No. JB06-F004 (10ML/bot)

375 mM Tris HCl, 50% Glycerol, 9% Betamercaptoethanol, 0.03% Bromophenol blue and 9% SDS at a pH of 6.8

T-Pro Laemmli (SDS sample) Reagent (non-reducing 6X)

No. JB06-F005 (10ML/bot)

375 mM Tris HCl, 50% Glycerol, 0.03% Bromophenol blue and 9% SDS at a pH of 6.8

T-Pro LDS Protein sample Reagent (reducing 4X)

No. JB06-F006 (10ML/bot)

988 mM Tris HCl, 2.04 mM EDTA, 40% Glycerol, 8% Betamercaptoethanol, 0.88mM Coomassie blue, 0.7 mM Phenol Red and 8% LDS at a pH of 8.5

T-Pro LDS Protein sample Reagent (non-reducing 4X)

No. JB06-F007 (10ML/bot)

988 mM Tris HCl, 2.04 mM EDTA, 40% Glycerol, 0.88mM Coomassie blue, 0.7 mM Phenol Red and 8% LDS at a pH of 8.5

Running Buffer

T-Pro Tris-Glycine-SDS running buffer (10X)

No. JB07-G001 (500ML/bot)

0.25 M Tris base, 1.92 M Glycine and 1% SDS at a pH8.3

T-Pro Tris-Glycine-Native running buffer (10X)

No. JB07-G002 (500ML/bot)

0.25 M Tris base, 1.92 M Glycine at a pH8.5

T-Pro MOPS/SDS running buffer (20X)

No. JB07-G003 (500ML/bot)

1 M MOPS, 1 M Tris base, 20 mM EDTA and 2% SDS at a pH7.7

Transfer Buffer

T-Pro Transfer Blotting buffer (10X)

No. JB08-H001 (500ML/bot)

0.25 M Tris base, 1.92 M Glycine

Just add methanol according to your protocol.

T-Pro Semi Dry Transfer Blotting buffer (10X)

No. JB08-H002 (500ML/bot)

0.48 M Tris base, 0.39 M Glycine and 0.375% SDS

Just add methanol according to your protocol.

T-Pro HQ Transfer Blotting buffer (20X)

No. JB08-H003 (500ML/bot)

For trans blot transfer

T-Pro HQ Transfer Blotting buffer (20X) 50 ml with deionized water 850 ml and add of methanol 100ml.

For semi dry transfer

T-Pro HQ Transfer Blotting buffer (20X) 50 ml with deionized water 400 ml and add of methanol 50ml.

Washing Buffer

T-Pro Phosphate-Buffered Saline (PBS, 10X)

No. JB09-I001 (500ML/bot)

100 mM Phosphate buffer, 27 mM KCl, 1.37 M NaCl at to pH7.4 after 1X dilution

T-Pro Tris-Buffered Saline (TBS, 10X)

No. JB09-I002 (500ML/bot)

0.5 M Tris HCl, 1.5 M NaCl at to pH7.4 after 1X dilution

T-Pro Washing buffer in PBS and Tween-20 (10X)

No. JB09-I003 (500ML/bot)

100 mM Phosphate buffer, 27 mM KCl, 1.37 M NaCl and 1% Tween-20 at to pH7.4 after 1X dilution

T-Pro Washing buffer in TBS and Tween-20 (10X)

No. JB09-I004 (500ML/bot)

0.5 M Tris HCl, 1.5 M NaCl and 1% Tween-20 at to pH7.4 after 1X dilution

T-Pro Western Blot Stripping Reagent

No. JB11-K002 (500ML/bot)

T-Pro Western Blot Stripping Reagent (EX)

No. JB11-K005 (500ML/bot)

  • Effective use of samples that are available in limited amounts.
  • Comparison of images obtained with different antibodies in the same blot.
  • Confirmation of results with the same or different antibodies.
  • It is simply more economical and less time consuming to reuse blots for re-probing and mass spectrometry.

Introduction:

Stripping Reagent for Western Blot is formulated to effectively remove the antibodies from Western blot that have been developed with chemiluminescence. The membrane can be nitrocellulose or PVDF/nylon. The stripped membrane can be re-probing for the Western Blot and for mass spectrometry.

Features

  • Strip blot in 5~15 minutes.
  • Incubation at room temperature.
  • No stench smelling and without addition of 2-mercaptoethanol or its analogs.
  • A propriety formulation with an eco-friendly and non-hazardous organic solvent (with light odor) and can completely disassociate the bound antibodies

Protocol of T-Pro Western Blot Stripping Reagent or (EX)

  • Pour 10 ml stripping reagent to a clean container and put the membrane in the container. Make sure that the membrane is fully submerged with the stripping buffer.
  • Incubate the membrane in stripping reagent at room temperature for 3 minutes with strong agitation for twice. For the high affinity antibodies, the incubation time need to be optimized, 5~10 minutes stripping at 37℃ is usually sufficient for most of antibodies.
  • Wash the membrane twice by using TBS-T or PBS-T at room temperature in large volumes (e.g. 100 ml) for 3 minutes.

Blocking Buffer

T-Pro Fast Blocking Buffer (in PBS or TBS)

(In PBS) No. JK92-W001 (500ML/bot)

(In TBS) No. JK92-W002 (500ML/bot)

T-Pro Protein Free Blocking Buffer (in PBS or TBS)

(In PBS) No. JK92-W003 (500ML/bot)

(In TBS) No. JK92-W004 (500ML/bot)

T-Pro Blocking Buffer

No. JK92-W005 (500ML/bot)

The T-Pro Blocking Buffers contain a proprietary compound for blocking excess binding sites in Western or ELISA blotting. This blocking buffer reduces or eliminates many of the problems encountered with traditional protein-blocking reagents, such as cross-reactivity and interference from glycosylation. Additionally, T-Pro Blocking Buffers are compatible with antibodies and avidin/biotin systems.

* Continue the blotting procedure do not using the T-Pro Blocking Buffer to dilute primary and secondary antibodies

  • The usage as described in these instructions may differ from other blocking solutions.
  • Use the blocking buffers at the supplied concentration; do not dilute blocking buffer.
  • A final concentration of 0.05% Tween-20 Detergent in the blocking buffer often improves blocking; however, it is not required nor recommended for all systems.
  • The protein-free blocking buffers may be used as a protein stabilizer for drying antigen- or antibody-coated microplates. Dry plate completely before sealing in a plastic bag with desiccant.

Stain solution

T-Pro Ponceau S Stain Solution 0.1% (1X)

No. JT90-R006M (500ML/bot)

T-Pro Ponceau S Stain Solution 0.5% (5X)

No. JT90-R007S (100ML/bot)

T-Pro Ponceau S Stain Solution is a ready‑to‑use membrane stain for evaluating protein transfer efficiency or for use in total protein normalization for western blot analysis. The stain has minimal nonspecific interactions with nitrocellulose or PVDF membranes, providing a reliable method for visualization of membrane‑bound proteins. Staining with Ponceau S Stain Solution results in pink-reddish protein bands that are easily photographed and reversed in less than 5 minutes. Subsequent immunodetection is unaffected, as the stain does not alter membrane‑bound proteins.

 

T-Pro Ponceau S Stain Solution is a ready‑to‑use membrane stain for evaluating protein transfer efficiency or for use in total protein normalization for western blot analysis. The stain has minimal nonspecific interactions with nitrocellulose or PVDF membranes, providing a reliable method for visualization of membrane‑bound proteins. Staining with Ponceau S Stain Solution results in pink-reddish protein bands that are easily photographed and reversed in less than 5 minutes. Subsequent immunodetection is unaffected, as the stain does not alter membrane‑bound proteins.