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antibody stripping buffer

antibody stripping buffer

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from the antigen and employs a lower pH to alter the structure of the antibody in such a way that the binding site is no longer active.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
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
StartingBlock

Serum and biotin-free single purified protein

  • Performs well with a wide range of antibodies and antibody combinations.
  • Compatible with streptavidin systems
  • Blocks in less than 15 minutes
  • Best for med-high abundant proteins or strong antibody affinity
  • High background with current blocking buffer
  • Stripping and reprobing western blots
PBS
TBS
PBST
TBST
Blocker FL

Single purified protein

  • Blocks excess nonspecific binding sites to help reduce background fluorescence,
  • Works with both nitrocellulose and low fluorescence PVDF
  • Detergent-free
  • Blocks in 15-30 minutes
  • Fluorescence western blotting
  • Imaging and storage of dry fluorescence blots
10X
Pierce Clear Milk Blocking Buffer

Clarified and stabilized milk proteins

  • High performance replacement for homemade milk blocking buffers in Western blotting applications
  • Long shelf-life at room temperature
  • Use when high background seen with Non-fat Milk
  • Fluorescent and chemiluminescent applications
Borate, pH 7.6
SuperBlock

Serum and biotin-free single purified glycoprotein

  • Protein-based formulation does not contain any immunoglobulins, albumin or endogenous biotin, making it compatible in many situations where traditional blocking agents fail.
  • Biotin-free for use with streptavidin system
  • Blocks in less than 10 minutes
  • High background with current blocking buffer
PBS
TBS
PBST
TBST
SEA BLOCK Blocking Buffer

Steelhead salmon serum

  • Useful in detection methods involving mammalian samples.
  • Particularly effective in applications involving fluorescence imaging
  • Mammalian samples
  • Fluorescence western blotting
PBS
Blocker BSA

Purified bovine serum albumin

  • 10% solutions of high-quality bovine serum albumin
  • Single purified protein provides fewer chances of cross-reaction with assay components than serum or milk solutions
  • Use when targeting phospho- proteins
  • Best to use when storing reused antibodies in blocker
PBS
TBS
Blocker Casein

Purified casein

  • Single-protein blocking buffer provides fewer chances of cross-reaction with assay components than serum or milk solutions
  • Use when high background seen with Non-fat milk blockers
PBS
TBS
Blocker BLOTTO

Non-fat dry milk

  • Ready-to-use 5% solution of nonfat powdered milk
  • Convenience- ready-to-use
  • More consistent product over home-made blockers
TBS
Protein-Free

Non-protein blocking compound

  • Minimizes or eliminates cross-reactivity associated with protein-based blocking buffers.
  • Sample-and-antibody combinations require the elimination of all possible exogenous animal proteins in the assay system to avoid cross-reaction or quenching of the desired probe function
  • Use when protein-based blockers cause high background
PBS
TBS
PBST
TBST
Pierce Fast Blocking Buffer

Single purified protein

  • Blocks in 5 minutes
  • When time is essential
TBS
SuperSignal Western Blot Enhancer

Membrane treatment for low abundance or poor immunoreactivity antibodies

  • Pre-treatment for nitrocellulose membranes
  • Reduces the amount of primary antibody required for probing
  • Use when primary antibody is 3 to 100-fold less primary antibody than is usually used to detect the protein of interest
Ready-to-use
antibody stripping buffer
antibody stripping buffer

 

Stripping and re-probing of Western blots offers several advantages:

  1. Conservation of samples that are expensive or available only in limited quantities;
  2. Analysis of a given blot using several different antibodies, e.g. subtype- or isoform-specific antibodies;
  3. Re-analysis of anomalous results and confirmation with the same or a different antibody;
  4. Correcting errors in incubation with the wrong antibody;
  5. 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

20-abx090656
  • EUR 189.00
  • EUR 230.00
  • 100 ml
  • 200 ml
  • Shipped within 5-10 working days.

WB / Antibody Stripping Buffer

abx090671-200ml 200 ml
EUR 189
  • Shipped within 5-10 working days.

West Ez Stripping Buffer

S2100-006 60ml
EUR 81

West Ez Stripping Buffer, Economic Size

S2100-050 500ml
EUR 212

West Ez Stripping Buffer, Economic Size

S2100-100 2x500ml
EUR 280

Western blot recycling stripping buffer, 50 ml (10X)

90101 50 ml
EUR 238

Phospho Antibody Stripping Solution

AKR-102 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

P5W3 NULL
EUR 0

T-Pro Western Blot Stripping Reagent

JB11-K002 500ml/BT
EUR 178

Antibody diluent buffer

AR1106-2 12ml
EUR 91

Fixation Buffer

22015 100mL
EUR 149
Description: Minimum order quantity: 1 unit of 100mL

Permeabilization Buffer

22016 100mL
EUR 149
Description: Minimum order quantity: 1 unit of 100mL

RIPA Buffer

2114-100
EUR 158

RIPA Buffer

2114-500
EUR 463

Dilution Buffer

1066-100
EUR 153

Dilution Buffer

1066-400
EUR 262

Wash Buffer

1210-200
EUR 196

Wash Buffer

abx098952-20ml 20 ml
EUR 91
  • Shipped within 5-7 working days.

Coating Buffer

abx098970-1vial 1 vial
EUR 154
  • Shipped within 5-7 working days.

Blocking Buffer

abx098972-1vial 1 vial
EUR 154
  • Shipped within 5-7 working days.

Lysis Buffer

abx098984-LysisBuffer120ml Lysis Buffer 1 (20 ml)
EUR 154
  • Shipped within 5-7 working days.

Lysis Buffer

abx098984-LysisBuffer3100ml Lysis Buffer 3 (100ml)
EUR 230
  • Shipped within 5-7 working days.

Lysis Buffer

abx098984-LysisBuffer420ml Lysis Buffer 4 (20 ml)
EUR 154
  • Shipped within 5-7 working days.

Tricine buffer

20-abx082551
  • EUR 217.00
  • EUR 189.00
  • 100 g
  • 25 g
  • Shipped within 5-10 working days.

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