Western Blot Stripping Buffer
Course of
- Warmth the buffer to 50°C
- Add the buffer to a small plastic area which has a good lid; use a amount which will cowl the membrane
- Add the membrane. Incubate at 50°C for as a lot as 45 min with some agitation
- Eradicate the reply as required for ß-mercaptoethanol based totally buffers
- Rinse the membrane beneath working water faucet for 1–2 min
- Traces of ß-mercaptoethanol will hurt the antibodies. Wash extensively for 5 min in TBST
- Ready for blocking
The Western BLoT Stripping Buffer is a solution for eradicating essential and secondary antibodies from probed Western blot membranes. Antibody elimination with this buffer can occur beneath mild circumstances (room temperature, 30 min incubation), minimizing lack of immobilized protein from the membrane. When using a PVDF membrane, the equivalent membrane is likely to be stripped and reprobed 2–5 events. After stripping, membranes is likely to be re-probed, each with a novel focus of essential antibody or with a completely completely totally different essential antibody.
Stripping is the time interval used to elucidate the elimination of essential and secondary antibodies from a western blot membrane. Stripping is useful when one must analysis a number of protein on the equivalent blot, as an illustration a protein of curiosity and a loading administration. When probing for a lot of targets, stripping and re-probing a single membrane instead of working and blotting a lot of gels has the good thing about saving samples, provides, and time.
It is not advisable to make quantitative comparisons of targets probed sooner than and after stripping given that course of removes some sample protein from the membrane. For the same trigger, a stripped membrane should not be probed to show the absence of a protein.
Thermo Scientific Restore Western Blot Stripping Buffer safely and efficiently removes essential and secondary antibodies from nitrocellulose and PVDF membranes to allow chemiluminescent Western blots to be reprobed.
Choices of Restore Western Blot Stripping Buffer:
• Saves time—no should re-run gels and blots
• Saves costly sample—re-probe the membrane using the equivalent aim sample
• Environment friendly—formulation is additional atmosphere pleasant at stripping antibodies than selfmade buffers
• Mild—would not hurt the aim antigen all through stripping allowing atmosphere pleasant reprobing
• Odor-free—no mercaptans means no acrid odors
• Economical—cheaper than totally different industrial stripping buffers
Product Particulars
Performing gel electrophoresis and duplicate immunoblot assays to test new essential antibodies or antibody concentrations is time-consuming and dear. Restore Western Blot Stripping Buffer eliminates this waste when detecting immunoblots with chemiluminescent Western blotting substrates. Restore Stripping Buffer provides clear and atmosphere pleasant elimination of essential and secondary antibodies from immunoblots with out eradicating or damaging the immobilized antigen allowing blots to be stripped and reprobed with confidence.
Chemiluminescent Western blot detection with reagents equal to Thermo Scientific SuperSignal Substrates for horseradish peroxidase is probably going one of many commonest and delicate methods in use instantly. Because of these substrates do not precipitate and bind to membrane surfaces, Western blots detected by chemiluminescence is likely to be stripped with reagents that take away affinity-bound essential and secondary antibodies. To be environment friendly, a stripping buffer should be sturdy adequate to disassociate sure antibodies nonetheless mild adequate to depart the transferred aim proteins intact on the nitrocellulose or PVDF membrane. Restore Western Blot Stripping Buffer has these traits.
By stripping and reprobing, there is no such thing as a such factor as a should waste unusual or costly samples by working a lot of gels with the intention to probe for numerous targets. A single membrane from one gel is likely to be stripped with Restore Western Blot Stripping Buffer to remove the primary antibodies. Stripping the blot takes solely 15 to 30 minutes, counting on the affinity of the primary antibody. After stripping, block and reprobe with a model new essential antibody. Alternatively, a blot is likely to be stripped and reprobed with adjusted antibody concentrations to optimize circumstances after buying initially poor outcomes.
Capabilities
• Reuse a nitrocellulose or PVDF blot to detect a novel aim with a novel essential antibody
• Reprobe a blot to applicable or optimize antibody concentrations which have been ineffective the first time
Protocol Summary
• Wash blot to remove chemiluminescent substrate.
• Incubate blot in Restore Western Blot Stripping Buffer for 5 to 15 minutes at 37°C (room temperature is sufficient for some antibodies).
• Take away blot and wash in Wash Buffer (TBS-T or PBS-T).
• Check out for sufficient elimination of antibodies.
• Perform subsequent immunoblot experiment.
 R - Jagged - 1(188 - 206), Notch Ligand (AA: Cys-Asp-Asp-Tyr-Tyr-Tyr-Gly-Phe-Gly-Cys-Asn-Lys-Phe-Gly-Arg-Pro-Arg-Asp-Asp) (MW: 2291.46)) R-JAG (188 - 206) R - Jagged - 1(188 - 206), Notch Ligand (AA: Cys-Asp-Asp-Tyr-Tyr-Tyr-Gly-Phe-Gly-Cys-Asn-Lys-Phe-Gly-Arg-Pro-Arg-Asp-Asp) (MW: 2291.46) |
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| SP-86688-1 | Alpha Diagnostics | 1 mg | EUR 164 |
) MCP-1 Monocyte Chemotactic Protein-1 Mouse Recombinant Protein (CCL2) |
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| PROTP10148-1 | BosterBio | Regular: 10ug | EUR 317 |
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Description: Monocyte Chemotactic Protein-1 Mouse Recombinant produced in E.Coli is a single,non-glycosylated, polypeptide chain containing 125 amino acids and having a molecular mass of 14 kDa. ;The MCP-1 is purified by proprietary chromatographic techniques. |
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) MCP-1 Monocyte Chemotactic Protein-1 Human Recombinant Protein (CCL2) |
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| PROTP13500-1 | BosterBio | Regular: 20ug | EUR 317 |
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Description: Monocyte Chemotactic Protein-1 Human Recombinant also known as Monocyte Chemotactic and Activating Factor (MCAF) produced in E.Coli is a non-glycosylated, Polypeptide chain containing 76 amino acids and having a molecular mass of 8.6kDa. ;The MCP-1 is purified by proprietary chromatographic techniques. |
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) Amyloid ?-Protein (1-15) |
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| A1003-1 | ApexBio | 1 mg | EUR 113 |
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Description: Beta-amyloid protein (A beta), a 39-43 amino acid peptide composed of a portion of the transmembrane domain and the extracellular domain of the amyloid precursor protein (APP), is also the principal component of amyloid. |
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, human recombinant protein) Neuregulin/Heregulin-1? (NRG-1?/HRG-1?), human recombinant protein |
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| P1054-1 | ApexBio | 1 mg | EUR 3947 |
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Description: Neuregulin (NRG) is a signaling protein for ErbB2/ErbB4 receptor heterodimers on the cardiac muscle cells and plays an important role in heart structure and function through inducing cardiomyocyte differentiation |
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 IL-1-beta Interleukin-1 betaHuman Recombinant Protein |
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| PROTP01584-1 | BosterBio | Regular: 10ug | EUR 317 |
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Description: Interleukin-1 beta Human Recombinant produced in E.Coli is a non-glycosylated, Polypeptide chain containing 153 amino acids and having a molecular mass of 17000 Dalton.;The IL-1b is purified by proprietary chromatographic techniques. |
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 MMP-1 Matrix Metalloproteinase-1 Human Recombinant Protein |
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| PROTP03956-1 | BosterBio | Regular: 20ug | EUR 317 |
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Description: MMP 1 Human Recombinant produced in E.coli is a single, non-glycosylated polypeptide chain containing 393 amino acids (100-469a.a) and having a molecular mass of 45kDa. MMP 1 is fused to a 23 amino acid His-tag at N-terminus. |
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 HIV-1 Tat Protein Peptide |
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| B1433-1 | ApexBio | 1 mg | EUR 128 |
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Description: HIV-1 Tat Protein Peptide |
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 ApoA-1, human recombinant protein |
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| P1052-.1 | ApexBio | 100 µg | EUR 313 |
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Description: Apolipoprotein A-1 (ApoA-1) is a glycoprotein produced in the liver and intestine that is the major protein component of high density lipoprotein (HDL) particles. ApoA-1 is involved in the reverse transport of cholesterol from peripheral tissues to the liver for recycling and excretion. |
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ApoA-1, human recombinant protein |
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| P1052-1 | ApexBio | 1 mg | EUR 1553 |
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Description: Apolipoprotein A-1 (ApoA-1) is a glycoprotein produced in the liver and intestine that is the major protein component of high density lipoprotein (HDL) particles. ApoA-1 is involved in the reverse transport of cholesterol from peripheral tissues to the liver for recycling and excretion. |
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 WNT-1, human recombinant protein |
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| P1068-1 | ApexBio | 1 mg | EUR 6940 |
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Description: The WNT gene family compose of structurally related genes that encode secreted signaling proteins. These proteins have been involved in oncogenesis and in several developmental processes, including regulation of cell fate and patterning during embryogenesis. |
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 Recombinant Human ANG-1 Protein |
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| PROTQ15389-1 | BosterBio | 20ug | EUR 317 |
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Description: Angiopoietin-1 (Ang-1) is a secreted ligand for Tie-2, a tyrosine-kinase receptor expressed primarily on vascular endothelial cells and early hematopoietic cells. Ang-1/ Tie-2 signaling promotes angiogenesis during the development, remodeling, and repair of the vascular system. Transgenic mice lacking expression of either Ang-1 or Tie-2 fail to develop a fully functional cardiovascular system and die before birth. Postnatally, the angiogenic activity of Ang-1/Tie-2 is required during normal tissue repair and remodeling of the female endometrium in the menstrual cycle. Ang-1/Tie-2 signaling appears to be regulated by Angiopoietin-2 (Ang-2), a natural antagonist for Tie-2 that exerts its effects through an internal autocrine loop mechanism. In addition to suppressing endothelial cell activation by inhibiting the expression of adhesion and inflammatory molecules, Ang-1 enhances endothelial cell survival and capillary morphogenesis, and lessens capillary permeability. As such, Ang-1 has a potential to become an effective therapeutic agent for treating various endothelium disorders, including several severe human pulmonary diseases. The efficacy of cell-based Ang-1 gene therapy for acute lung injury (ALI) has recently been studied in a rat model of ALI (1). The results of this study show that such therapy can markedly improve lung condition and suggest that Ang-1 therapy may represent a potential new strategy for the treatment and/or prevention of acute respiratory distress injury (ARDI), a significant cause of morbidity and mortality in critically ill patients. Recombinant human ANG-1, derived from HeLa cells, is a C-terminal histidine tagged glycoprotein which migrates with an apparent molecular mass of 60.0 – 70.0 kDa by SDS-PAGE under reducing conditions. Sequencing analysis shows N-terminal sequences starting with Ser-20 and with Asp-70 of the 498 amino acid precursor protein. |
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 Recombinant Murine Cardiotrophin-1 Protein |
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| PROTQ60753-1 | BosterBio | 10ug | EUR 317 |
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Description: CT-1 is a member of the IL-6 family of cytokines which also includes LIF, CNTF, OSM (Oncostatin M), IL-11, IL-6 and possibly NNT-1/BCSF-3. CT-1 is a pleiotropic cytokine which is expressed in various tissues including the adult heart, skeletal muscle, ovary, colon, prostate and fetal lung and signals through the LIF receptor and the gp130 receptor subunit. CT-1 has the ability to induce cardiac myocyte hypertrophy, and enhances the survival of cardiomyocyte and different neuronal populations. Recombinant murine Cardiotrophin-1 is a 21.3 kDa protein consisting of 202 amino acid residues. |
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 Recombinant Human Gremlin-1 Protein |
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| PROTO60565-1 | BosterBio | 50ug | EUR 317 |
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Description: Gremlin-1 (isoform-1) belongs to a group of diffusible proteins which bind to ligands of the TGF-β family and regulate their activity by inhibiting their access to signaling receptors. The interplay between TGF-β ligands and their natural antagonists has major biological significance during development processes, in which cellular response can vary considerably depending upon the local concentration of the signaling molecule. Gremlin is highly expressed in the small intestine, fetal brain, and colon and lower expression in brain, prostate, pancreas and skeletal muscle. Gremlin-1 regulates multiple functions in early development by specifically binding to and inhibiting the function of BMP-2, -4, and -7. It also plays a role in carcinogenesis and kidney branching morphogenesis. Recombinant Gremlin-1 is a 18.3 kDa protein containing 160 amino acid residues. |
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 ORM1 Orosomucoid 1 Human protein |
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| PROTP02763-1 | BosterBio | Regular: 10ug | EUR 317 |
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Description: The Human Orosomucoid 1 produced from Human pooled serum has a molecular mass of 21.56kDa (calculated without glycosylation) containing 183 amino acid residues. |
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 Recombinant Human Galectin-1 Protein |
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| PROTP09382-1 | BosterBio | 50ug | EUR 317 |
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Description: Lectins, of either plant or animal origin, are carbohydrate binding proteins that interact with glycoprotein and glycolipids on the surface of animal cells. The Galectins are lectins that recognize and interact with β-galactoside moieties. Galectin-1 is an animal lectin that has been shown to interact with CD3, CD4, and CD45. It induces apoptosis of activated T-cells and T-leukemia cell lines and inhibits the protein phosphatase activity of CD45. Recombinant human Galectin-1 is a 14.5 kDa protein containing 134 amino acid residues. |
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 Recombinant Human PECAM-1 Protein |
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| PROTP16284-1 | BosterBio | 50ug | EUR 317 |
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Description: PECAM is transmembrane glycoprotein that belongs to the Ig-related superfamily of adhesion molecules. It is highly expressed at endothelial cell junctions, and also expressed in platelets and in most leukocyte sub-types. The primary function of PECAM-1 is the mediation of leukocyte-endothelial cell adhesion and signal transduction. PECAM-1 has been implicated in the pathogenesis of various inflammation related disorders, including thrombosis, multiple sclerosis (MS), and rheumatoid arthritis. The human PECAM-1 gene codes for a 738 amino acid transmembrane glycoprotein containing a 118 amino acid cytoplasmic domain, a 19 amino acid transmembrane domain, and a 574 amino acid extracellular domain. Recombinant human PECAM-1 is a 572 amino acid glycoprotein comprising the extracellular domain of PECAM-1. Monomeric glycosylated PECAM-1 migrates at an apparent molecular weight of approximately 80.0-95.0 kDa by SDS-PAGE analysis under reducing conditions |
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 Canine IGF-1 Recombinant Protein |
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| R00148-1 | BosterBio | 5ug/vial | EUR 259 |
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Description: Insulin-like growth factor 1 (IGF-1) is a primary mediator of the effects of growth hormone (GH) and has growth-promoting effects on almost every cell in the body. IGF-1 can also regulate cell growth and development, especially in nerve cells, as well as cellular DNA synthesis. Canine IGF-1 Recombinant Protein is purified insulin-like growth factor 1 (IGF-1) produced in yeast. |
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 RAD51D (1-328 a.a.) Human Recombinant Protein) (1-328) RAD51D (1-328 a.a.) Human Recombinant Protein |
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| PROTO75771-1 | BosterBio | Regular: 10ug | EUR 317 |
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Description: RAD51D (1-328) Human Recombinant produced in E. coli is. a single polypeptide chain containing 351 amino acids and having a molecular mass of 37.4kDa. RAD51D (1-328) is fused to a 23 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques. |
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 VSNL1 Visinin-Like Protein-1 Human Recombinant Protein |
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| PROTP62760-1 | BosterBio | Regular: 50ug | EUR 317 |
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Description: VSNL1 Human Recombinant produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 191 amino acids (1-191 a.a.) and having a molecular mass of 22.1kDa.;The VSNL1 is purified by proprietary chromatographic techniques. |
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 DKK1 Dickkopf-Related Protein 1 Human Recombinant Protein |
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| PROTO94907-1 | BosterBio | Regular: 20ug | EUR 317 |
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Description: DKK1 Human Recombinant produced in E.Coli is a single, non-glycosylated polypeptide chain containing 258 amino acids (32-266 a.a) and having a molecular mass of 28.2kDa.;DKK1 is fused to a 23 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques. |
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 AssayMax ELISA Kit) Human KRAB-associated Protein 1 (KAP-1) AssayMax ELISA Kit |
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| EK2802-1 | AssayPro | 96 Well Plate | EUR 477 |
 GAD1 iso1 Glutamate Decarboxylase 1 Isoform-1 Human Recombinant Protein |
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| PROTQ99259-1 | BosterBio | Regular: 20ug | EUR 317 |
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Description: GAD1 iso1 Human Recombinant produced in E.Coli is a single, non-glycosylated polypeptide chain containing 617 amino acids (1-594 a.a) and having a molecular mass of 69.3kDa. GAD1 iso1 is fused to a 23 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques. |
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 Envelope-1, Dengue Virus Subtype 1 Recombinant Protein, Insect Cells |
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| PROTB1PNJ5-1 | BosterBio | Regular: 10ug | EUR 317 |
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Description: Recombinant Dengue Virus Subtype 1 produced in Insect Cells is a polypeptide chain containing amino acids 281-675 and having a molecular weight of approximately 50kDa. ;Dengue Envelope-1 is purified by proprietary chromatographic technique. |
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 TGF-b-1 Transforming Growth Factor-beta 1 Human protein |
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| PROTP01137-1 | BosterBio | Regular: 2.5ug | EUR 1157 |
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Description: Human Transforming Growth Factor-beta 1 purified from Human Platelets having a molecular mass of 25kDa.;The TGF-b 1 is purified by proprietary chromatographic techniques. |
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