Name: Antibody Stripping Solution
Catalog Number: L7710A, 500ml* GV0480, 500ml 5X
* Enough reagent for 3000 cm2 or 200-400 membrane strips or 20-40 standard blots
• Storage: Store at RT (for long storage: +4C°) (L)
No pungent smelling mercaptoethanol saves time
Antibody stripping is performed at room temperature. No need to heat the blots, saves costly sample
Strip antibodies economical in only 15 minutes at room temperature
Blots can be avoided by re-blocking in most cases.
Western Blot Stripping Buffer
Western BLOT stripping buffer is a solution to remove primary and secondary antibodies from the examined western blot membranes. Antibody removal with this buffer can occur under mild conditions (room temperature, 30 min incubation), minimizing the loss of immobilized proteins from the membrane. When using PVDF membranes, the same membrane can be stripped and reprobed 2-5 times. After stripping, the membrane can be re-probed, either with a different concentration of primary antibody or with a completely different primary antibody.
Several protocols have been published for antibody stripping from western blots, including low pH, heat and detergents, and the use of chaotropic agents. Three recommended protocols are presented below. The first applies to any chemiluminescent substrate system and uses a combination of detergent and heat to release the antibody. The second is commonly used for applications where the antibody has to be separated from the antigen and employs a lower pH to alter the structure of the antibody in such a way that the binding site is no longer active.
The third protocol uses the ReBlot™Plus Western Blot Recycling Kit that has been specifically designed to isolate antibodies from western blot membranes. Benefits of this approach include
Cat. # | Product | Size | License | Quantity | Details | |
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T7135A | Western BLoT Stripping Buffer | 500 mL | * | |||
The Western BLoT Stripping Buffer is a reagent that can remove primary and secondary antibodies from Western blot membranes. After treatment with the Stripping Buffer, the membrane can be reused; it is possible to probe the membrane with either a different concentration of primary antibody or with an entirely different primary antibody. With this product, the antibody removal reaction proceeds under relatively mild conditions (room temperature, 30 minutes), and therefore there is very little loss of immobilized protein from the membrane. When using a PVDF membrane, the same membrane can be stripped and re-probed 2–5 times. |
Blocking buffer | Blocking agent | Highlights | When to use | Available formats |
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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 |

Stripping and re-probing of Western blots offers several advantages:
- Conservation of samples that are expensive or available only in limited quantities;
- Analysis of a given blot using several different antibodies, e.g. subtype- or isoform-specific antibodies;
- Re-analysis of anomalous results and confirmation with the same or a different antibody;
- Correcting errors in incubation with the wrong antibody;
- Cost savings in reagents and time by reusing the same blot.
Neither of these methods will remove the colored precipitate resulting from chromogenic detection systems (eg, BCIP, 4CN, DAB and TMB). However, it is still possible to analyze the blot with another antibody specific for a different target protein.
Typically, these protocols should be used for qualitative purposes only, until it is established that stripping does not quantitatively affect a given antigen. Depending on the method and type of membrane used, many antigens will withstand at least 5 stripping cycles. However, it should be kept in mind that small portions of membrane-stabilized proteins will be removed during each stripping cycle. When multiple antigens are detected sequentially, it is recommended to start with antigens that are expected to be in low abundance or to give a low signal.
Here are some additional recommendations when planning a Western blot experiment with one or more rounds of antibody stripping.
- PVDF membranes are more robust than nitrocellulose and are therefore recommended for any protocol involving antibody stripping.
- Drying of PVDF membranes immediately after transfer from the SDS-PAGE gel improves binding of proteins to the membrane and is particularly recommended when multiple stripping is planned. Dry PVDF membranes must be rewetted with alcohol prior to the first round of immunodetection.
- Detect low-abundance antigens first.
- Use low-affinity antibodies before high-affinity antibodies.
- Important: Although drying of a PVDF blot is recommended immediately after transfer, the blot should not be allowed to dry between rounds of immunodetection. Any residual antibody molecules will bind permanently to the membrane if it is allowed to dry.
WB / Antibody Stripping Buffer |
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abx090671-200ml | Abbexa | 200 ml | EUR 189 |
West Ez Stripping Buffer |
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S2100-006 | GenDepot | 60ml | EUR 81 |
West Ez Stripping Buffer, Economic Size |
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S2100-050 | GenDepot | 500ml | EUR 212 |
West Ez Stripping Buffer, Economic Size |
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S2100-100 | GenDepot | 2x500ml | EUR 280 |
Western blot recycling stripping buffer, 50 ml (10X) |
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90101 | Alpha Diagnostics | 50 ml | EUR 238 |
Phospho Antibody Stripping Solution |
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AKR-102 | Cell Biolabs | 1 kit | EUR 392 |
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 |
T-Pro Western Blot Stripping Reagent |
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JB11-K002 | T-Pro Biotechnology | 500ml/BT | EUR 178 |
Antibody diluent buffer |
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AR1106-2 | BosterBio | 12ml | EUR 91 |
PBS Buffer |
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RM00012 | Abclonal | 2L | EUR 102 |
10xTaq Buffer |
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PCRB60 | Bio Basic | 4x1.5ml, 6ml | EUR 58.7 |
Wash Buffer |
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KF17356 | Neuromics | 500 ml | EUR 179 |
Fixation Buffer |
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22015 | Biotium | 100mL | EUR 149 |
Description: Minimum order quantity: 1 unit of 100mL |
Permeabilization Buffer |
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22016 | Biotium | 100mL | EUR 149 |
Description: Minimum order quantity: 1 unit of 100mL |
ADA buffer |
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AD0003 | Bio Basic | 25g | EUR 64.79 |
Wash Buffer |
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abx098952-20ml | Abbexa | 20 ml | EUR 91 |
Coating Buffer |
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abx098970-1vial | Abbexa | 1 vial | EUR 154 |
Blocking Buffer |
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abx098972-1vial | Abbexa | 1 vial | EUR 154 |
Lysis Buffer |
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abx098984-LysisBuffer120ml | Abbexa | Lysis Buffer 1 (20 ml) | EUR 154 |
Lysis Buffer |
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abx098984-LysisBuffer3100ml | Abbexa | Lysis Buffer 3 (100ml) | EUR 230 |
Lysis Buffer |
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abx098984-LysisBuffer420ml | Abbexa | Lysis Buffer 4 (20 ml) | EUR 154 |
Tricine buffer |
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20-abx082551 | Abbexa |
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Binding Buffer |
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abx290019-50ml | Abbexa | 50 ml | EUR 230 |
Wash Buffer |
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abx293002-30ml | Abbexa | 30 ml | EUR 105 |