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A quantitative western blot method utilizing TMB

• Lisa
• July 21, 2021May 25, 2021
• 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
A12001	EpiGentek	EUR 232.26 EUR 117.70	350 µg 350 ug

Goat Anti-Rat Secondary Antibody, Biotin Conjugated
A12002	EpiGentek	EUR 232.26 EUR 117.70	350 µg 350 ug

Anti-Galectin 1 Antibody
A00470-2	BosterBio	100ug/vial	EUR 352.8

Anti-Galectin-1 antibody
PAab03314	Lifescience Market	100 ug	EUR 426

anti- Galectin-1 antibody
FNab03314	FN Test	100µg	EUR 606.3
Description: Antibody raised against Galectin-1

anti- Galectin-1 antibody
FNab03315	FN Test	100µg	EUR 658.5
Description: Antibody raised against Galectin-1

Anti-Galectin-1 antibody
STJ93201	St John's Laboratory	200 µl	EUR 236.4
Description: Rabbit polyclonal to Galectin-1.

Anti-Galectin-1 antibody
STJ98716	St John's Laboratory	200 µl	EUR 236.4
Description: Rabbit polyclonal to Galectin-1.

Anti-Galectin 1 (1A8)
YF-MA13984	Abfrontier	100 ug	EUR 435.6
Description: Mouse monoclonal to Galectin 1

Human Galectin-1 (LGALS1)
1-CSB-EP012882HU	Cusabio	EUR 456.00 EUR 256.80 EUR 1570.80 EUR 672.00 EUR 1047.60 EUR 314.40	100ug 10ug 1MG 200ug 500ug 50ug
Description: Recombinant Human Galectin-1(LGALS1) expressed in E.coli

Human Galectin-1 (LGALS1)
1-CSB-EP012882HUb1	Cusabio	EUR 456.00 EUR 256.80 EUR 1570.80 EUR 672.00 EUR 1047.60 EUR 314.40	100ug 10ug 1MG 200ug 500ug 50ug
Description: Recombinant Human Galectin-1(LGALS1) expressed in E.coli

Human Genomic DNA
X11000	EpiGentek	Ask for price EUR 235.40	0.2 ml 0.2 ml

Anti-Galectin 1/Lgals1 Antibody
A00470	BosterBio	100ug/vial	EUR 400.8

Anti-Galectin 1 Biotinylated Antibody
A00470-Biotin	BosterBio	50ug/vial	EUR 352.8

anti-Galectin 1 (1E8-1B2)
LF-MA10174	Abfrontier	100 ug	EUR 435.6
Description: Mouse monoclonal to Galectin 1

Anti-Galectin 1/LGALS1 Antibody
PA1422	BosterBio	100ug/vial	EUR 400.8

Anti-Galectin 1/LGALS1 Antibody
PB9240	BosterBio	100ug/vial	EUR 400.8

Anti-galectin-1 (mouse) antibody
STJ72519	St John's Laboratory	100 µg	EUR 430.8

Human Brain Genomic DNA
X11001	EpiGentek	Ask for price EUR 77.00	10 µg 10 ul

Anti-Galectin-13 (GAL13) / Placental Protein 13 (PP13) Monoclonal Antibody
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
GST-ANTI-2	Detroit R&D	50 uL	EUR 336

Polyclonal Goat anti-GST p-form
GST-ANTI-3	Detroit R&D	50 uL	EUR 336

LGALS8 Human, Galectin-8 Human Recombinant Protein, His Tag
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
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
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
EG3311-1	AssayPro	96 Well Plate	EUR 572.4

Human Galectin-4 (Gal-4) AssayMax ELISA Kit
EG3312-1	AssayPro	96 Well Plate	EUR 572.4

anti-Galectin 3
YF-PA24079	Abfrontier	50 ul	EUR 400.8
Description: Mouse polyclonal to Galectin 3

anti-galectin 9
YF-PA24081	Abfrontier	50 ul	EUR 400.8
Description: Mouse polyclonal to galectin 9

anti-Galectin 3
YF-PA12946	Abfrontier	50 ug	EUR 435.6
Description: Mouse polyclonal to Galectin 3

anti-Galectin 3
YF-PA12947	Abfrontier	100 ul	EUR 483.6
Description: Rabbit polyclonal to Galectin 3

anti-Galectin 3
YF-PA12948	Abfrontier	100 ug	EUR 483.6
Description: Rabbit polyclonal to Galectin 3

anti-Galectin 8
YF-PA12951	Abfrontier	50 ul	EUR 435.6
Description: Mouse polyclonal to Galectin 8

anti-Galectin 13
YF-PA18585	Abfrontier	50 ug	EUR 435.6
Description: Mouse polyclonal to Galectin 13

Human Galectin-1(LGALS1) ELISA kit
1-CSB-EL012882HU	Cusabio	EUR 964.80 EUR 6118.80 EUR 3244.80	1 plate of 96 wells 10 plates of 96 wells each 5 plates of 96 wells each
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
7-00427	CHI Scientific	10µg	Ask for price

Recombinant Human Galectin-1
7-00428	CHI Scientific	50µg	Ask for price

Recombinant Human Galectin-1
7-00429	CHI Scientific	1mg	Ask for price

Human Galectin 1 Protein
20-abx066725	Abbexa	EUR 744.00 EUR 326.40 EUR 2230.80 EUR 878.40 EUR 543.60	100 ug 10 ug 1 mg 200 ug 50 ug

Galectin-1, human recombinant
4648-10	Biovision	each	EUR 170.4

Galectin-1, human recombinant
4648-1000	Biovision	each	EUR 1977.6

Galectin-1, human recombinant
4648-50	Biovision	each	EUR 314.4

Anti-human Galectin-3 antibody
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)
1-CSB-EP012882BO	Cusabio	EUR 733.20 EUR 370.80 EUR 2192.40 EUR 1126.80 EUR 1461.60 EUR 476.40	100ug 10ug 1MG 200ug 500ug 50ug
Description: Recombinant Bovine Galectin-1(LGALS1) expressed in E.coli

SARS-CoV-2 Spike S1 RBD Protein, Human Fc-Fusion, Avi-Tag
E80025	EpiGentek	EUR 635.80 EUR 3934.70	100 ul 1 ml

Anti-Galectin-9 Antibody
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
PAab03310	Lifescience Market	100 ug	EUR 494.4

Anti-Galectin 4 antibody
PAab03311	Lifescience Market	100 ug	EUR 426

Anti-Galectin 8 antibody
PAab03312	Lifescience Market	100 ug	EUR 426

Anti-Galectin 9 antibody
PAab03313	Lifescience Market	100 ug	EUR 426

Anti-Galectin-3 antibody
PAab03316	Lifescience Market	100 ug	EUR 426

Anti-Galectin-7 antibody
PAab03318	Lifescience Market	100 ug	EUR 426

anti-Galectin-3 (6G2)
LF-MA20336	Abfrontier	100 ug	EUR 424.8
Description: Mouse monoclonal to Galectin-3

anti- Galectin 2 antibody
FNab03310	FN Test	100µg	EUR 702
Description: Antibody raised against Galectin 2

anti- Galectin 4 antibody
FNab03311	FN Test	100µg	EUR 606.3
Description: Antibody raised against Galectin 4

anti- Galectin 8 antibody
FNab03312	FN Test	100µg	EUR 606.3
Description: Antibody raised against Galectin 8

anti- Galectin 9 antibody
FNab03313	FN Test	100µg	EUR 606.3
Description: Antibody raised against Galectin 9

anti- Galectin-3 antibody
FNab03316	FN Test	100µg	EUR 606.3
Description: Antibody raised against Galectin-3

anti- Galectin-3 antibody
FNab03317	FN Test	100µg	EUR 658.5
Description: Antibody raised against Galectin-3

anti- Galectin-7 antibody
FNab03318	FN Test	100µg	EUR 606.3
Description: Antibody raised against Galectin-7

Anti-Galectin-2 antibody
STJ93202	St John's Laboratory	200 µl	EUR 236.4
Description: Rabbit polyclonal to Galectin-2.

Anti-Galectin-4 antibody
STJ93203	St John's Laboratory	200 µl	EUR 236.4
Description: Rabbit polyclonal to Galectin-4.

Anti-Galectin-7 antibody
STJ93204	St John's Laboratory	200 µl	EUR 236.4
Description: Rabbit polyclonal to Galectin-7.

Anti-Galectin-8 antibody
STJ93205	St John's Laboratory	200 µl	EUR 236.4
Description: Rabbit polyclonal to Galectin-8.

Anti-Galectin-9 antibody
STJ93206	St John's Laboratory	200 µl	EUR 236.4
Description: Rabbit polyclonal to Galectin-9.

Anti-Galectin-3 antibody
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
STJ97549	St John's Laboratory	200 µl	EUR 236.4
Description: Mouse monoclonal to Galectin-3 (6B8).

Anti-Galectin-3 antibody
STJ97550	St John's Laboratory	200 µl	EUR 236.4
Description: Mouse monoclonal to Galectin-3 (8D7).

Anti-Galectin-3 antibody
STJ97551	St John's Laboratory	200 µl	EUR 236.4
Description: Mouse monoclonal to Galectin-3 (5D9).

Anti-Galectin-3 antibody
STJ97601	St John's Laboratory	200 µl	EUR 236.4
Description: Mouse monoclonal to Galectin-3 (2F9).

Anti-Galectin-3 antibody
STJ97604	St John's Laboratory	200 µl	EUR 236.4
Description: Mouse monoclonal to Galectin-3 (16E6).

Anti-Galectin-3 antibody
STJ97605	St John's Laboratory	200 µl	EUR 236.4
Description: Mouse monoclonal to Galectin-3 (1H4).

Anti-Galectin-3 antibody
STJ97715	St John's Laboratory	200 µl	EUR 236.4
Description: Mouse monoclonal to Galectin-3.

Anti-Galectin-3 antibody
STJ16100369	St John's Laboratory	1 mL	EUR 1256.4

Anti-Galectin-3 antibody
STJ16100782	St John's Laboratory	100 µg	EUR 864

Anti-Galectin-3 antibody
STJ180106	St John's Laboratory	0.1 ml	EUR 254.4

Anti-Galectin 3 antibody
STJ190057	St John's Laboratory	200 µl	EUR 236.4
Description: Unconjugated Mouse monoclonal to Galectin 3 (AS1A24)

Anti-Galectin 3 antibody
STJ73098	St John's Laboratory	100 µg	EUR 430.8

Anti-Galectin 8 (3E5)
YF-MA13986	Abfrontier	100 ug	EUR 435.6
Description: Mouse monoclonal to Galectin 8

Galectin 1 antibody
70R-11939	Fitzgerald	100 ug	EUR 483.6
Description: Rabbit polyclonal Galectin 1 antibody

Galectin 1 antibody
70R-49997	Fitzgerald	100 ul	EUR 292.8
Description: Purified Polyclonal Galectin 1 antibody

Galectin 1 antibody
70R-13963	Fitzgerald	100 ug	EUR 386.4
Description: Affinity purified Rabbit polyclonal Galectin 1 antibody

Galectin 1 protein
30R-1385	Fitzgerald	100 ug	EUR 268.8
Description: Purified recombinant Human Galectin 1 protein

Galectin 1 protein
30R-2371	Fitzgerald	50 ug	EUR 423.6
Description: Purified recombinant Human Galectin 1 protein

Galectin 1 Antibody
49673-100ul	SAB	100ul	EUR 399.6

Galectin 1 Antibody
49673-50ul	SAB	50ul	EUR 286.8

Galectin 1 protein
30R-AG003	Fitzgerald	10 ug	EUR 159.6
Description: Purified recombinant Human Galectin 1 protein

Galectin-1 Protein
20-abx263505	Abbexa	EUR 276.00 EUR 2148.00 EUR 393.60	10 ug 1 mg 50 ug

Galectin-1 Protein
20-abx261634	Abbexa	EUR 4101.60 EUR 393.60 EUR 276.00	1 mg 20 ug 5 ug

Galectin 1 antibody
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.