
A quantitative western blot method utilizing TMB
Lisa
- 0
A quantitative western blot technique using TMB: Comparability with the usual technique
Fairly a number of molecular natural experiments carried out all by way of the world require the detection or quantification of a protein of curiosity. Western blotting is among the many hottest methods used for this purpose and offers quantitative knowledge with the assistance of specialized software program program. However, its dependence on the picture that is captured, and the background and the absence of a typical protocol forestall the tactic from being completely quantitative. To beat these obstacles, we present a straightforward and reliable assay that is very similar to the frequent technique, excluding the ultimate stage of band visualization and quantification.
We recommend that small objects of the blot that embody the protein of curiosity could be decrease and dipped in a small amount of three,3′,5,5′-Tetramethylbenzidine (TMB) decision, giving a colorimetric signal with linear dependence on the quantity of the protein. The response is stopped with H2 SO4 , and the signal is measured in a plate reader. This modification displays extreme linearity with out additional costs and could be utilized for every purified proteins and proteins current in a lysate. The outcomes obtained with our proposed technique have been in distinction with these obtained by the usual approach and proved to be further reliable. This textual content is protected by copyright. All rights reserved.

Goat Anti-Rabbit Secondary Antibody, Biotin Conjugated |
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A12001 | EpiGentek |
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Goat Anti-Rat Secondary Antibody, Biotin Conjugated |
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A12002 | EpiGentek |
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Anti-Galectin 1 Antibody |
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A00470-2 | BosterBio | 100ug/vial | EUR 352.8 |
Anti-Galectin-1 antibody |
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PAab03314 | Lifescience Market | 100 ug | EUR 426 |
anti- Galectin-1 antibody |
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FNab03314 | FN Test | 100µg | EUR 606.3 |
Description: Antibody raised against Galectin-1 |
anti- Galectin-1 antibody |
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FNab03315 | FN Test | 100µg | EUR 658.5 |
Description: Antibody raised against Galectin-1 |
Anti-Galectin-1 antibody |
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STJ93201 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Rabbit polyclonal to Galectin-1. |
Anti-Galectin-1 antibody |
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STJ98716 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Rabbit polyclonal to Galectin-1. |
Anti-Galectin 1 (1A8) |
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YF-MA13984 | Abfrontier | 100 ug | EUR 435.6 |
Description: Mouse monoclonal to Galectin 1 |
Human Galectin-1 (LGALS1) |
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1-CSB-EP012882HU | Cusabio |
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Description: Recombinant Human Galectin-1(LGALS1) expressed in E.coli |
Human Galectin-1 (LGALS1) |
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1-CSB-EP012882HUb1 | Cusabio |
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Description: Recombinant Human Galectin-1(LGALS1) expressed in E.coli |
Human Genomic DNA |
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X11000 | EpiGentek |
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Anti-Galectin 1/Lgals1 Antibody |
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A00470 | BosterBio | 100ug/vial | EUR 400.8 |
Anti-Galectin 1 Biotinylated Antibody |
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A00470-Biotin | BosterBio | 50ug/vial | EUR 352.8 |
anti-Galectin 1 (1E8-1B2) |
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LF-MA10174 | Abfrontier | 100 ug | EUR 435.6 |
Description: Mouse monoclonal to Galectin 1 |
Anti-Galectin 1/LGALS1 Antibody |
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PA1422 | BosterBio | 100ug/vial | EUR 400.8 |
Anti-Galectin 1/LGALS1 Antibody |
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PB9240 | BosterBio | 100ug/vial | EUR 400.8 |
Anti-galectin-1 (mouse) antibody |
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STJ72519 | St John's Laboratory | 100 µg | EUR 430.8 |
Human Brain Genomic DNA |
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X11001 | EpiGentek |
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Anti-Galectin-13 (GAL13) / Placental Protein 13 (PP13) Monoclonal Antibody |
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M08143-1 | BosterBio | 100ug/vial | EUR 476.4 |
Description: Mouse Monoclonal Galectin-13 (GAL13) / Placental Protein 13 (PP13) Antibody. Validated in IHC and tested in Human. |
Polyclonal Goat anti-GST μ-form |
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GST-ANTI-2 | Detroit R&D | 50 uL | EUR 336 |
Polyclonal Goat anti-GST p-form |
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GST-ANTI-3 | Detroit R&D | 50 uL | EUR 336 |
LGALS8 Human, Galectin-8 Human Recombinant Protein, His Tag |
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PROTO00214-1 | BosterBio | Regular: 10ug | EUR 380.4 |
Description: LGALS8 Human Recombinant produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 337 amino acids (1-317 a.a.) and having a molecular mass of 37.9 kDa. The LGALS8 is fused to a 20 amino acid His-Tag at N-terminus and purified by proprietary chromatographic techniques. |
LGALS3 Human, Galectin-3 Human Recombinant Protein, His Tag |
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PROTP17931-1 | BosterBio | Regular: 25ug | EUR 380.4 |
Description: LGALS3 Human Recombinant produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 270 amino acids (1-250 a.a.) and having a molecular mass of 28.3 kDa._x000D_ The LGALS3 is fused to a 20 amino acid His-Tag at N-terminus and purified by proprietary chromatographic techniques. _x000D_ |
LGALS7 Human, Galectin-7 Human Recombinant Protein, His Tag |
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PROTP47929-1 | BosterBio | Regular: 20ug | EUR 380.4 |
Description: Galectin-7 Human Recombinant fused with a 20 amino acid His tag at N-terminus produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 156 amino acids (1-136 a.a.) and having a molecular mass of 17.2kDa. ;The Galectin-7 is purified by proprietary chromatographic techniques. |
Human Galectin-3 (Gal-3) AssayMax ELISA Kit |
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EG3311-1 | AssayPro | 96 Well Plate | EUR 572.4 |
Human Galectin-4 (Gal-4) AssayMax ELISA Kit |
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EG3312-1 | AssayPro | 96 Well Plate | EUR 572.4 |
anti-Galectin 3 |
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YF-PA24079 | Abfrontier | 50 ul | EUR 400.8 |
Description: Mouse polyclonal to Galectin 3 |
anti-galectin 9 |
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YF-PA24081 | Abfrontier | 50 ul | EUR 400.8 |
Description: Mouse polyclonal to galectin 9 |
anti-Galectin 3 |
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YF-PA12946 | Abfrontier | 50 ug | EUR 435.6 |
Description: Mouse polyclonal to Galectin 3 |
anti-Galectin 3 |
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YF-PA12947 | Abfrontier | 100 ul | EUR 483.6 |
Description: Rabbit polyclonal to Galectin 3 |
anti-Galectin 3 |
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YF-PA12948 | Abfrontier | 100 ug | EUR 483.6 |
Description: Rabbit polyclonal to Galectin 3 |
anti-Galectin 8 |
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YF-PA12951 | Abfrontier | 50 ul | EUR 435.6 |
Description: Mouse polyclonal to Galectin 8 |
anti-Galectin 13 |
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YF-PA18585 | Abfrontier | 50 ug | EUR 435.6 |
Description: Mouse polyclonal to Galectin 13 |
Human Galectin-1(LGALS1) ELISA kit |
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1-CSB-EL012882HU | Cusabio |
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Description: Quantitativesandwich ELISA kit for measuring Human Galectin-1(LGALS1) in samples from serum, plasma, tissue homogenates, cell lysates. Now available in a cost efficient pack of 5 plates of 96 wells each, conveniently packed along with the other reagents in 5 separate kits. |
Recombinant Human Galectin-1 |
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7-00427 | CHI Scientific | 10µg | Ask for price |
Recombinant Human Galectin-1 |
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7-00428 | CHI Scientific | 50µg | Ask for price |
Recombinant Human Galectin-1 |
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7-00429 | CHI Scientific | 1mg | Ask for price |
Human Galectin 1 Protein |
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20-abx066725 | Abbexa |
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Galectin-1, human recombinant |
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4648-10 | Biovision | each | EUR 170.4 |
Galectin-1, human recombinant |
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4648-1000 | Biovision | each | EUR 1977.6 |
Galectin-1, human recombinant |
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4648-50 | Biovision | each | EUR 314.4 |
Anti-human Galectin-3 antibody |
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STJ15100163 | St John's Laboratory | 250 µg | EUR 403.2 |
Description: This monoclonal antibody is for studies of antigen expression in cells and tissue sections using immunocytochemistry and immunoprecipitation |
Bovine Galectin-1 (LGALS1) |
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1-CSB-EP012882BO | Cusabio |
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Description: Recombinant Bovine Galectin-1(LGALS1) expressed in E.coli |
SARS-CoV-2 Spike S1 RBD Protein, Human Fc-Fusion, Avi-Tag |
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E80025 | EpiGentek |
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Anti-Galectin-9 Antibody |
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A03415 | BosterBio | 100ul | EUR 476.4 |
Description: Rabbit Polyclonal Antibody for Galectin-9 Antibody (LGALS9) detection.tested for WB in Human, Mouse, Rat. |
Anti-Galectin 2 antibody |
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PAab03310 | Lifescience Market | 100 ug | EUR 494.4 |
Anti-Galectin 4 antibody |
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PAab03311 | Lifescience Market | 100 ug | EUR 426 |
Anti-Galectin 8 antibody |
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PAab03312 | Lifescience Market | 100 ug | EUR 426 |
Anti-Galectin 9 antibody |
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PAab03313 | Lifescience Market | 100 ug | EUR 426 |
Anti-Galectin-3 antibody |
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PAab03316 | Lifescience Market | 100 ug | EUR 426 |
Anti-Galectin-7 antibody |
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PAab03318 | Lifescience Market | 100 ug | EUR 426 |
anti-Galectin-3 (6G2) |
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LF-MA20336 | Abfrontier | 100 ug | EUR 424.8 |
Description: Mouse monoclonal to Galectin-3 |
anti- Galectin 2 antibody |
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FNab03310 | FN Test | 100µg | EUR 702 |
Description: Antibody raised against Galectin 2 |
anti- Galectin 4 antibody |
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FNab03311 | FN Test | 100µg | EUR 606.3 |
Description: Antibody raised against Galectin 4 |
anti- Galectin 8 antibody |
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FNab03312 | FN Test | 100µg | EUR 606.3 |
Description: Antibody raised against Galectin 8 |
anti- Galectin 9 antibody |
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FNab03313 | FN Test | 100µg | EUR 606.3 |
Description: Antibody raised against Galectin 9 |
anti- Galectin-3 antibody |
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FNab03316 | FN Test | 100µg | EUR 606.3 |
Description: Antibody raised against Galectin-3 |
anti- Galectin-3 antibody |
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FNab03317 | FN Test | 100µg | EUR 658.5 |
Description: Antibody raised against Galectin-3 |
anti- Galectin-7 antibody |
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FNab03318 | FN Test | 100µg | EUR 606.3 |
Description: Antibody raised against Galectin-7 |
Anti-Galectin-2 antibody |
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STJ93202 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Rabbit polyclonal to Galectin-2. |
Anti-Galectin-4 antibody |
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STJ93203 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Rabbit polyclonal to Galectin-4. |
Anti-Galectin-7 antibody |
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STJ93204 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Rabbit polyclonal to Galectin-7. |
Anti-Galectin-8 antibody |
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STJ93205 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Rabbit polyclonal to Galectin-8. |
Anti-Galectin-9 antibody |
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STJ93206 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Rabbit polyclonal to Galectin-9. |
Anti-Galectin-3 antibody |
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STJ96945 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Galectin-3 is a protein encoded by the LGALS3 gene which is approximately 26,1 kDa. Galectin-3 is localised to the cytoplasm and nucleus. It is involved in NF-kappaB signalling, advanced glycosylation end product receptor signalling and cell adhesion. It is a galactose-specific lectin which binds IgE and may mediate the stimulation by CSPG4 of endothelial cells migration along with the alpha-3, beta-1 integrin. It is characterized by an N-terminal proline-rich tandem repeat domain and a single C-terminal carbohydrate recognition domain. Galectin-3 is expressed mainly in the colonic epithelium and is also abundantly expressed in activated macrophages. Mutations in the LGALS3 gene may result in follicular adenoma and thyroid cancer. STJ96945 was developed from clone 6G2 and was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen. This antibody detects endogenous galectin-3 proteins. |
Anti-Galectin-3 antibody |
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STJ97549 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Mouse monoclonal to Galectin-3 (6B8). |
Anti-Galectin-3 antibody |
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STJ97550 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Mouse monoclonal to Galectin-3 (8D7). |
Anti-Galectin-3 antibody |
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STJ97551 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Mouse monoclonal to Galectin-3 (5D9). |
Anti-Galectin-3 antibody |
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STJ97601 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Mouse monoclonal to Galectin-3 (2F9). |
Anti-Galectin-3 antibody |
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STJ97604 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Mouse monoclonal to Galectin-3 (16E6). |
Anti-Galectin-3 antibody |
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STJ97605 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Mouse monoclonal to Galectin-3 (1H4). |
Anti-Galectin-3 antibody |
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STJ97715 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Mouse monoclonal to Galectin-3. |
Anti-Galectin-3 antibody |
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STJ16100369 | St John's Laboratory | 1 mL | EUR 1256.4 |
Anti-Galectin-3 antibody |
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STJ16100782 | St John's Laboratory | 100 µg | EUR 864 |
Anti-Galectin-3 antibody |
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STJ180106 | St John's Laboratory | 0.1 ml | EUR 254.4 |
Anti-Galectin 3 antibody |
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STJ190057 | St John's Laboratory | 200 µl | EUR 236.4 |
Description: Unconjugated Mouse monoclonal to Galectin 3 (AS1A24) |
Anti-Galectin 3 antibody |
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STJ73098 | St John's Laboratory | 100 µg | EUR 430.8 |
Anti-Galectin 8 (3E5) |
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YF-MA13986 | Abfrontier | 100 ug | EUR 435.6 |
Description: Mouse monoclonal to Galectin 8 |
Galectin 1 antibody |
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70R-11939 | Fitzgerald | 100 ug | EUR 483.6 |
Description: Rabbit polyclonal Galectin 1 antibody |
Galectin 1 antibody |
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70R-49997 | Fitzgerald | 100 ul | EUR 292.8 |
Description: Purified Polyclonal Galectin 1 antibody |
Galectin 1 antibody |
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70R-13963 | Fitzgerald | 100 ug | EUR 386.4 |
Description: Affinity purified Rabbit polyclonal Galectin 1 antibody |
Galectin 1 protein |
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30R-1385 | Fitzgerald | 100 ug | EUR 268.8 |
Description: Purified recombinant Human Galectin 1 protein |
Galectin 1 protein |
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30R-2371 | Fitzgerald | 50 ug | EUR 423.6 |
Description: Purified recombinant Human Galectin 1 protein |
Galectin 1 Antibody |
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49673-100ul | SAB | 100ul | EUR 399.6 |
Galectin 1 Antibody |
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49673-50ul | SAB | 50ul | EUR 286.8 |
Galectin 1 protein |
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30R-AG003 | Fitzgerald | 10 ug | EUR 159.6 |
Description: Purified recombinant Human Galectin 1 protein |
Galectin-1 Protein |
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20-abx263505 | Abbexa |
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Galectin-1 Protein |
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20-abx261634 | Abbexa |
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Galectin 1 antibody |
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70R-GR002 | Fitzgerald | 100 ug | EUR 571.2 |
Description: Affinity purified Rabbit polyclonal Galectin 1 antibody |
Western Blot Analysis in Ewing Sarcoma
Western blot is an experimental approach used to analysis protein expression. In Ewing sarcoma, as in numerous totally different diseases, Western blot provides particulars in regards to the extent of protein expression in quite a few cell circumstances, in contrast with totally different tissues or upon induced molecular changes.
Based on the specific pattern of protein expression of the tissue, along with on the traits of the protein of curiosity, the antibodies and protocol of Western blot may be modified consistent with utterly totally different specs. Proper right here we describe a number of of these peculiarities in physique of Ewing sarcoma topic.
Identification of novel fibronectin-binding proteins by 2D-far Western blot in atypical enteropathogenic Escherichia coli serotype O55:H7
Atypical enteropathogenic Escherichia coli (aEPEC) is a subgroup of EPEC, which is among the many principal pathogens answerable for lethal diarrhoea in kids. In distinction with typical EPEC (tEPEC), aEPEC lack an EAF (EPEC adherence concern) plasmid (pEAF), which encodes a group of virulence-associated genes. The extracellular matrix (ECM) aspect of human cells has been reported to be an important facet throughout the interaction between host and bacterial pathogens. On this evaluation, a 2D-Far Western blot approach was carried out to identifiy the bacterial proteins that will bind to fibronectin, most likely probably the most widespread constituents of ECM. A whole of 17 protein spots have been acknowledged, along with 4 outer membrane proteins (OMPs), notably, OmpC, OmpD, OmpX and LamB. In vitro analysis have been used to search out out whether or not or not these OMPs have been involved throughout the adherence course of.
By the use of indirect immunofluorescence assays, Four OMPs could very effectively be seen on the surfaces of host cells. After incubating the cells with the recombinant proteins, the adhesion cost of the O55:H7 isolate was decreased. Furthermore, the deletion of OmpX and LamB could decrease the adhesion cost of WT. Taken collectively, a high-throughput screening approach for host ECM-binding proteins based mostly totally on 2D Far-Western blot was established, and Four outer membrane proteins acknowledged by this system have been found to be involved throughout the adherence course of.
Validation of superior reference genes for qRT-PCR and Western blot analyses in marine Emiliania huxleyi – virus model system
Objectives: To hunt for a set of reference genes for reliable gene expression analysis throughout the globally important marine coccolithophore Emiliania huxleyi -virus model system.
Methods and outcomes: Fifteen housekeeping genes (CDKA, CYP15, EFG3, POLAI, RPL30, RPL13, SAMS, COX1, GPB1-2, HSP90, TUA, TUB, UBA1, CAM3 and GAPDH) have been evaluated for his or her stability as potential reference genes for qRT-PCR using ΔCt, geNorm, NormFinder, Bestkeeper and RefFinder software program program. CDKA, TUA and TUB genes have been examined as loading controls for Western blot within the similar sample panel. Furthermore, purpose genes associated to cell apoptosis, i.e., metacaspase genes, have been utilized to validate the variety of reference genes. The analysis outcomes demonstrated that putative housekeeping genes exhibited necessary variations in every mRNA and protein content material materials all through virus an an infection. After a whole analysis with all of the algorithms, CDKA and GAPDH have been helpful as most likely probably the most regular reference genes for E huxleyi virus (EhV) an an infection cures. For Western blot, necessary variation was seen for TUA and TUB, whereas CDKA was stably expressed, in line with the outcomes of qRT-PCR.
Conclusions: CDKA and GAPDH are the best choice for gene and protein expression analysis than the alternative candidate reference genes under EhV an an infection circumstances.
Significance and affect of the look at: The regular interior administration genes acknowledged on this work will help to reinforce the accuracy and reliability of gene expression analysis and obtain notion into difficult E. huxleyi-EhV interaction regulatory networks.
Western Blot Analysis in Ewing Sarcoma
Western blot is an experimental approach used to analysis protein expression. In Ewing sarcoma, as in numerous totally different diseases, Western blot provides particulars in regards to the extent of protein expression in quite a few cell circumstances, in contrast with totally different tissues or upon induced molecular changes.
Based on the specific pattern of protein expression of the tissue, along with on the traits of the protein of curiosity, the antibodies and protocol of Western blot may be modified in accordance to utterly totally different specs. Proper right here we describe a number of of those peculiarities in physique of Ewing sarcoma topic.
Identification of novel fibronectin-binding proteins by 2D-far Western blot in atypical enteropathogenic Escherichia coli serotype O55:H7
Atypical enteropathogenic Escherichia coli (aEPEC) is a subgroup of EPEC, which is among the many principal pathogens answerable for lethal diarrhoea in kids. In distinction with typical EPEC (tEPEC), aEPEC lack an EAF (EPEC adherence concern) plasmid (pEAF), which encodes a group of virulence-associated genes. The extracellular matrix (ECM) aspect of human cells has been reported to be an important facet throughout the interaction between host and bacterial pathogens.
On this evaluation, a 2D-Far Western blot approach was carried out to identifiy the bacterial proteins that will bind to fibronectin, most likely probably the most widespread constituents of ECM. A whole of 17 protein spots have been acknowledged, along with 4 outer membrane proteins (OMPs), notably, OmpC, OmpD, OmpX and LamB. In vitro analysis have been used to search out out whether or not or not these OMPs have been involved throughout the adherence course of. By the use of indirect immunofluorescence assays, Four OMPs could very effectively be seen on the surfaces of host cells. After incubating the cells with the recombinant proteins, the adhesion cost of the O55:H7 isolate was decreased.
Furthermore, the deletion of OmpX and LamB can moreover decrease the adhesion cost of WT. Taken collectively, a high-throughput screening approach for host ECM-binding proteins based mostly totally on 2D Far-Western blot was established, and Four outer membrane proteins acknowledged by this system have been found to be involved throughout the adherence course of.