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Western Blot Stripping Buffer

The Western BLoT Stripping Buffer is an answer for eradicating main and secondary antibodies from probed Western blot membranes. Antibody elimination with this buffer can happen underneath gentle circumstances (room temperature, 30 min incubation), minimizing lack of immobilized protein from the membrane. When utilizing a PVDF membrane, the identical membrane might be stripped and reprobed 2–5 occasions. After stripping, membranes might be re-probed, both with a unique focus of main antibody or with a wholly totally different main antibody.

Stripping is the time period used to explain the elimination of main and secondary antibodies from a western blot membrane. Stripping is beneficial when one needs to research multiple protein on the identical blot, as an illustration a protein of curiosity and a loading management. When probing for a number of targets, stripping and re-probing a single membrane as a substitute of working and blotting a number of gels has the benefit of saving samples, supplies, and time.

It isn’t advisable to make quantitative comparisons of targets probed earlier than and after stripping for the reason that process removes some pattern protein from the membrane. For a similar cause, a stripped membrane shouldn’t be probed to display the absence of a protein.

Process

  1. Heat the buffer to 50°C
  2. Add the buffer to a small plastic field which has a decent lid; use a quantity that may cowl the membrane
  3. Add the membrane. Incubate at 50°C for as much as 45 min with some agitation
  4. Eliminate the answer as required for ß-mercaptoethanol primarily based buffers
  5. Rinse the membrane underneath working water faucet for 1–2 min
  6. Traces of ß-mercaptoethanol will harm the antibodies. Wash extensively for five min in TBST
  7. Prepared for blocking

 

Thermo Scientific Restore Western Blot Stripping Buffer safely and successfully removes main and secondary antibodies from nitrocellulose and PVDF membranes to permit chemiluminescent Western blots to be reprobed.

Options of Restore Western Blot Stripping Buffer:

• Saves time—no have to re-run gels and blots
• Saves expensive pattern—re-probe the membrane utilizing the identical goal pattern
• Efficient—formulation is extra environment friendly at stripping antibodies than selfmade buffers
• Light—doesn’t harm the goal antigen throughout stripping permitting environment friendly reprobing
• Odor-free—no mercaptans means no acrid odors
• Economical—cheaper than different industrial stripping buffers

Product Particulars
Performing gel electrophoresis and duplicate immunoblot assays to check new main antibodies or antibody concentrations is time-consuming and costly. Restore Western Blot Stripping Buffer eliminates this waste when detecting immunoblots with chemiluminescent Western blotting substrates. Restore Stripping Buffer supplies clear and environment friendly elimination of main and secondary antibodies from immunoblots with out eradicating or damaging the immobilized antigen permitting blots to be stripped and reprobed with confidence.

Chemiluminescent Western blot detection with reagents equivalent to Thermo Scientific SuperSignal Substrates for horseradish peroxidase is likely one of the commonest and delicate strategies in use immediately. As a result of these substrates don’t precipitate and bind to membrane surfaces, Western blots detected by chemiluminescence might be stripped with reagents that take away affinity-bound main and secondary antibodies. To be efficient, a stripping buffer have to be sturdy sufficient to disassociate certain antibodies however light sufficient to depart the transferred goal 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 have to waste uncommon or expensive samples by working a number of gels with the intention to probe for various targets. A single membrane from one gel might be stripped with Restore Western Blot Stripping Buffer to take away the first antibodies. Stripping the blot takes solely 15 to 30 minutes, relying on the affinity of the first antibody. After stripping, block and reprobe with a brand new main antibody. Alternatively, a blot might be stripped and reprobed with adjusted antibody concentrations to optimize circumstances after acquiring initially poor outcomes.

 

antibody stripping buffer
antibody stripping buffer



Functions
• Reuse a nitrocellulose or PVDF blot to detect a unique goal with a unique main antibody
• Reprobe a blot to appropriate or optimize antibody concentrations that have been ineffective the primary time

Protocol Abstract
• Wash blot to take away chemiluminescent substrate.
• Incubate blot in Restore Western Blot Stripping Buffer for five to 15 minutes at 37°C (room temperature is enough for some antibodies).
• Take away blot and wash in Wash Buffer (TBS-T or PBS-T).
• Take a look at for enough elimination of antibodies.
• Carry out subsequent immunoblot experiment.

 

 

Amyloid ?-Protein (1-15)

A1003-1 1 mg
EUR 113
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.

Neuregulin/Heregulin-1? (NRG-1?/HRG-1?), human recombinant protein

P1054-1 1 mg
EUR 3947
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

IL-1-beta Interleukin-1 betaHuman Recombinant Protein

PROTP01584-1 Regular: 10ug
EUR 317
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.

MMP-1 Matrix Metalloproteinase-1 Human Recombinant Protein

PROTP03956-1 Regular: 20ug
EUR 317
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.

ORM1 Orosomucoid 1 Human protein

PROTP02763-1 Regular: 10ug
EUR 317
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.

Recombinant Human Galectin-1 Protein

PROTP09382-1 50ug
EUR 317
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.

Recombinant Human Gremlin-1 Protein

PROTO60565-1 50ug
EUR 317
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.

Canine IGF-1 Recombinant Protein

R00148-1 5ug/vial
EUR 259
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.

Recombinant Human PECAM-1 Protein

PROTP16284-1 50ug
EUR 317
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

Recombinant Human ANG-1 Protein

PROTQ15389-1 20ug
EUR 317
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.

Recombinant Murine Cardiotrophin-1 Protein

PROTQ60753-1 10ug
EUR 317
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.

HIV-1 Tat Protein Peptide

B1433-1 1 mg
EUR 128
Description: HIV-1 Tat Protein Peptide

ApoA-1, human recombinant protein

P1052-.1 100 µg
EUR 313
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.

ApoA-1, human recombinant protein

P1052-1 1 mg
EUR 1553
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.

WNT-1, human recombinant protein

P1068-1 1 mg
EUR 6940
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.

(1-328) RAD51D (1-328 a.a.) Human Recombinant Protein

PROTO75771-1 Regular: 10ug
EUR 317
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.

DKK1 Dickkopf-Related Protein 1 Human Recombinant Protein

PROTO94907-1 Regular: 20ug
EUR 317
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.

VSNL1 Visinin-Like Protein-1 Human Recombinant Protein

PROTP62760-1 Regular: 50ug
EUR 317
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.

TGF-b-1 Transforming Growth Factor-beta 1 Human protein

PROTP01137-1 Regular: 2.5ug
EUR 1157
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.

Envelope-1, Dengue Virus Subtype 1 Recombinant Protein, Insect Cells

PROTB1PNJ5-1 Regular: 10ug
EUR 317
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.

Human KRAB-associated Protein 1 (KAP-1) AssayMax ELISA Kit

EK2802-1 96 Well Plate
EUR 477

GAD1 iso1 Glutamate Decarboxylase 1 Isoform-1 Human Recombinant Protein

PROTQ99259-1 Regular: 20ug
EUR 317
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.

Recombinant Human sTRAIL Receptor-1 Protein

PROTO00220-1 50ug
EUR 317
Description: TRAIL Receptor-1/DR4 and TRAIL Receptor-2/DR5 belong to the TNFR superfamily of transmembrane proteins and contain a cytoplasmic "death domain, " which can activate the cell's apoptotic machinery. These receptors are activated by binding to either membrane anchored or soluble TRAIL/Apo2L. Recombinant human soluble TRAIL Receptor-1/DR4 is a 22.7 kDa protein (215 amino acid residues) consisting of the TNFR homologous, cysteine rich portion of the extracellular domain.

NNT1 Neurotrophin-1 Human Recombinant Protein

PROTQ9UBD9-1 Regular: 10ug
EUR 317
Description: Neurotrophin-1 Human Recombinant (28-225) produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 199 amino acids and having a molecular mass of 22kDa.;The NNT-1 is purified by proprietary chromatographic techniques.

HPSE Heparanase-1 Human Recombinant Protein

PROTQ9Y251-1 Regular: 20ug
EUR 317
Description: HPSE Human Recombinant produced in E.Coli is a single, non-glycosylated polypeptide chain containing 531 amino acids (36-543 a.a) and having a molecular mass of 60kDa.;HPSE is fused to a 23 amino acid His-tag at N-terminus.

Canine IL-1 beta Recombinant Protein

R00101-1 5ug/vial
EUR 259
Description: IL-1 beta (IL-1β) is a member of the interleukin 1 family of cytokines. The IL-1 beta cytokine is produced by activated macrophages as a proprotein, which is proteolytically processed to its active form by caspase 1 (CASP1/ICE). This cytokine is an important mediator of the inflammatory response, and is involved in a variety of cellular activities, including cell proliferation, differentiation, and apoptosis. Canine IL-1 beta Recombinant Protein is purified interleukin-1 beta cytokine produced in yeast.

Canine IL-1 alpha Recombinant Protein

R01144-1 5ug/vial
EUR 259
Description: IL-1 alpha (IL-1α, IL-1F1) is a member of the interleukin 1 family of cytokines. IL-1 alpha is an inflammatory cytokine active in the initiation of the inflammatory reaction and in driving Th1 and Th17 inflammatory responses. Canine IL-1 alpha Recombinant Protein is purified interleukin-1 alpha cytokine produced in yeast.

Equine IFN beta 1 Recombinant Protein

R02041-1 5ug/vial
EUR 259
Description: IFN beta is a mammalian Type I inferferon, functionig as a regulator of cellular activity by interacting with cell-surface receptors and activating various signaling pathways. IFN beta produces antiviral, antibacterial, and anticancer properties. Equine IFN beta Recombinant Protein is purified IFN beta produced in yeast.