Stripping and re-probing of Western blots provides an a variety of benefits:
- Conservation of samples which is likely to be pricey or obtainable solely in restricted parts;
- Analysis of a given blot using numerous completely completely different antibodies, e.g. subtype- or isoform-specific antibodies;
- Re-analysis of anomalous outcomes and affirmation with the similar or a particular antibody;
- Correcting errors in incubation with the inaccurate antibody;
- Worth monetary financial savings in reagents and time by reusing the similar blot.
Western Blot Stripping Buffer
The Western BLoT Stripping Buffer is a solution for eradicating main and secondary antibodies from probed Western blot membranes. Antibody eradicating with this buffer can occur beneath delicate conditions (room temperature, 30 min incubation), minimizing lack of immobilized protein from the membrane. When using a PVDF membrane, the similar membrane could possibly be stripped and reprobed 2–5 situations. After stripping, membranes could possibly be re-probed, each with a particular focus of main antibody or with a very completely completely different main antibody.
Cat. # | Product | Measurement | License | Quantity | Particulars | |
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T7135A | Western BLoT Stripping Buffer | 500 mL | * | |||
The Western BLoT Stripping Buffer is a reagent which will take away main and secondary antibodies from Western blot membranes. After treatment with the Stripping Buffer, the membrane could possibly be reused; it is attainable to probe the membrane with each a particular focus of main antibody or with a very completely completely different main antibody. With this product, the antibody eradicating response proceeds beneath comparatively delicate conditions (room temperature, 30 minutes), and as a consequence of this reality there could also be little or no lack of immobilized protein from the membrane. When using a PVDF membrane, the similar membrane could possibly be stripped and re-probed 2–5 situations. |
Title: Antibody Stripping Decision
Catalog amount: L7710A, 500ml* GV0480, 500ml 5X
*Enough reagents for 3000 cm2 or 200-400 membrane strips or 20-40 commonplace blots
• Storage: Retailer at RT ( for a protracted storage: +4C°) (L)
● No pungent smelling mercaptoethanol Save time
● Antibody stripping is accomplished at room temperature. No heating of blots required Saves costly sample
● Strip antibodies in almost 15 minutes at room temperature Economical
● Reblocking of blots may be prevented in most conditions

Blocking buffer | Blocking agent | Highlights | When to utilize | On the market codecs |
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StartingBlock |
Serum and biotin-free single purified protein |
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PBS TBS PBST TBST |
Blocker FL |
Single purified protein |
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10X |
Pierce Clear Milk Blocking Buffer |
Clarified and stabilized milk proteins |
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Borate, pH 7.6 |
SuperBlock |
Serum and biotin-free single purified glycoprotein |
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PBS TBS PBST TBST |
SEA BLOCK Blocking Buffer |
Steelhead salmon serum |
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PBS |
Blocker BSA |
Purified bovine serum albumin |
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PBS TBS |
Blocker Casein |
Purified casein |
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PBS TBS |
Blocker BLOTTO |
Non-fat dry milk |
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TBS |
Protein-Free |
Non-protein blocking compound |
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PBS TBS PBST TBST |
Pierce Fast Blocking Buffer |
Single purified protein |
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TBS |
SuperSignal Western Blot Enhancer |
Membrane treatment for low abundance or poor immunoreactivity antibodies |
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Ready-to-use |
Plenty of protocols for antibody stripping from Western blots have been printed, along with these which benefit from low pH, heat and detergent, and chaotropic brokers. Three advisable protocols are provided below. The first is related to any chemiluminescent substrate system and makes use of a mix of detergent and heat to launch the antibodies. The second is commonly used for features the place antibodies should be separated from an antigen and employs low pH to vary the development of the antibody in such a signifies that the binding site is no longer energetic.
The third protocol employs the ReBlot™ Plus Western Blot Recycling Package deal which has been notably formulated for stripping antibodies from Western blot membranes. Benefits of this technique embody
- Avoidance of odor related to β-mercaptoethanol.
- Room temperature processing.
- Elimination of antibodies in 15 minutes.
None of these methods will take away the colored precipitates generated from chromogenic detection methods (e.g., BCIP, 4CN, DAB and TMB). Nonetheless, it is nonetheless attainable to analysis the blot with one different antibody explicit to a particular purpose protein.
Usually, these protocols should solely be used for qualitative features, besides it has been established that stripping does not quantitatively impact a given antigen. Relying upon the tactic and form of membrane used, many antigens will stand as much as not lower than 5 stripping cycles. Nonetheless, it is to be thought of that in each stripping cycle small elements of membrane-immobilized proteins is likely to be eradicated. When numerous antigens are to be detected sequentially, it’s strongly advisable to start with antigens which are anticipated to occur at lower abundance or yield a lot much less signal.
Listed beneath are some additional solutions when planning a Western blot experiment with numerous rounds of antibody stripping.
- PVDF membranes are additional sturdy than nitrocellulose and are as a consequence of this reality advisable for any protocol involving antibody stripping.
- Drying of PVDF membranes immediately after swap from the SDS-PAGE gel improves binding of proteins to the membrane and is particularly advisable when numerous stripping is deliberate. Dry PVDF membranes must be rewetted with alcohol earlier to the first spherical of immunodetection.
- Detect low-abundance antigens first.
- Use low-affinity antibodies sooner than high-affinity antibodies.
- Mandatory: Although drying of a PVDF blot is advisable immediately after swap, the blot should not be allowed to dry between rounds of immunodetection. Any residual antibody molecules will bind utterly to the membrane whether or not it’s allowed to dry.
Stripping Buffer |
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GR103020 | Genorise Scientific | 500 mL | EUR 89 |
WB Stripping Buffer |
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AR0153 | BosterBio | 100mL (for 10 assays for an 5 × 8.5cm blot) | EUR 133.2 |
West Ez Stripping Buffer |
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S2100-006 | GenDepot | 60ml | EUR 97.2 |
West Ez Stripping Buffer, Economic Size |
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S2100-050 | GenDepot | 500ml | EUR 254.4 |
West Ez Stripping Buffer, Economic Size |
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S2100-100 | GenDepot | 2x500ml | EUR 336 |
Western blot recycling stripping buffer, 50 ml (10X) |
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90101 | Alpha Diagnostics | 50 ml | EUR 285.6 |
Phospho Antibody Stripping Solution |
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AKR-102 | Cell Biolabs | 1 kit | EUR 470.4 |
Description: This reagent allows you to strip phospho antibodies from protein blots and subsequently re-probe the same blot. |
101Bio WB Stripping Solution |
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P5W3 | 101Bio | - | Ask for price |
Dura Mater Stripping Forcep Angled |
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INS6134 | Scientific Laboratory Supplies | EACH | EUR 112.8 |
Dura Mater Stripping Forcep Straight |
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INS6132 | Scientific Laboratory Supplies | EACH | EUR 112.8 |
T-Pro Western Blot Stripping Reagent |
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JB11-K002 | T-Pro Biotechnology | 500ml/BT | EUR 213.6 |
Human IgG-RF stripper/Adsorbent (1 ml, sufficient for stripping 50 samples of 100 ul human serum/plasma) |
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RFS-1 | Alpha Diagnostics | 1 ml | EUR 123.6 |
100ML ProtoLift Western Strip Buffer |
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NAT1256 | Scientific Laboratory Supplies | 100ML | EUR 92.7 |
PhastGel SDS Buffer Strips pack of 10 |
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17051601 | Scientific Laboratory Supplies | EACH | EUR 116.28 |
PhastGel Native Buffer Strips pack of 10 |
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17051701 | Scientific Laboratory Supplies | EACH | EUR 116.28 |
Antibody diluent buffer |
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AR1106-2 | BosterBio | 12ml | EUR 109.2 |
ExcelGel SDS Buffer Strips anode and cathode 6 pairs |
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17134201 | Scientific Laboratory Supplies | EACH | EUR 154.8 |
Corning 3D Clear Starter Kit - Includes Reagent/Antibody Buffer/Blocking Buffer/Penetration Buffer/Washing Buffers |
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5730 | Scientific Laboratory Supplies | EACH | EUR 955.2 |
Corning 3D Clear Antibody Buffer - 100mL |
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5735 | Scientific Laboratory Supplies | EACH | EUR 265.2 |
Corning 3D Clear Antibody Buffer - 30mL |
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5734 | Scientific Laboratory Supplies | EACH | EUR 134.4 |
ChonBlock Detection Antibody Dilution Buffer, 100 ml |
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90681 | Chondrex | 100 ml | EUR 155.7 |
Description: ChonBlock Detection Antibody Dilution Buffer |
Antibody Dilution Buffer for ICC and IHC |
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SF40010 | Neuromics | 10 ml | EUR 146.4 |
Western Blot Primary Antibody Diluent Buffer |
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abx090689-100ml | Abbexa | 100 ml | EUR 218.4 |
Borax Buffer 0.00996M |
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BOR0091 | Scientific Laboratory Supplies | 1L | EUR 76.8 |
TT Buffer (Tris-Tricine buffer) Primix powder |
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TD8133 | Bio Basic | 1PK, 10L | EUR 91.32 |
ADA buffer |
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AD0003 | Bio Basic | 25g | EUR 77.75 |
PBS Buffer |
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RM00012 | Abclonal | 2L | EUR 122.4 |
West-Ezier Antibody Dilution Buffer, ready-to-use |
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W3872-050 | GenDepot | 500ml | EUR 198 |