Avidin biotinylated enzyme complex

Manufactured by Vector Laboratories

Sourced in United States

The avidin-biotinylated enzyme complex is a versatile tool used in various biochemical and immunological applications. It consists of the protein avidin, which has a high affinity for the small molecule biotin. The biotinylated enzyme, when combined with the avidin, forms a stable complex that can be utilized for detecting or capturing target analytes in samples.

Automatically generated - may contain errors

Lab products found in correlation

3 3 diaminobenzidine (dab), by Vector Laboratories (3 mentions) Vector sg, by Vector Laboratories (2 mentions) Mab1281, by Merck Group (1 mentions) Diaminobenzidine substrate chromogen system, by Agilent Technologies (1 mentions) Ab14106, by Abcam (1 mentions) Ab94744, by Abcam (1 mentions) Dp260, by Olympus (1 mentions) Dako serum free blocking solution, by Agilent Technologies (1 mentions) 3 3 diaminobenzidine (dab), by Merck Group (1 mentions) Biotinylated secondary antibody, by Vector Laboratories (1 mentions) 3 3 diaminobenzidine substrate, by Vector Laboratories (1 mentions) Fluka eukitt quick hardening mounting medium, by Merck Group (1 mentions) P stat3, by Cell Signaling Technology (1 mentions) Automated quantitative pathology imaging system, by PerkinElmer (1 mentions) Nf κb p65, by Abcam (1 mentions) Permount, by Merck Group (1 mentions) Antigen unmasking solution, by Agilent Technologies (1 mentions) Biotinylated anti rabbit secondary antibody, by Vector Laboratories (1 mentions) Immpact novared peroxidase substrate, by Vector Laboratories (1 mentions) Tyrosine hydroxylase, by Merck Group (1 mentions)

Spelling variants of this product

Avidin/Biotinylated enzyme complex (ABC; (5 citations) Avidin/Biotinylated Enzyme Complex kit (2 citations) Avidin-biotinylated enzyme complex (ABC reagent, (2 citations) Avidin-biotinylated peroxidase enzyme complex (ABC)-based kit (2 citations)

8 protocols using avidin biotinylated enzyme complex

Immunohistochemistry for Neuronal Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Free-floating sections were blocked in 3% normal serum, then incubated in primary antibody with 1% serum overnight at RT. Tissue was incubated in secondary antibody (1:200) with 1% serum for 3 h, then immersed in an avidin-biotinylated enzyme complex (Vector Laboratories) for 2 h, then either stained with DAB or Vector SG (Vector Laboratories).
The primary antibodies used in either bright-field or fluorescent immunohistochemistry were tyrosine hydroxylase (TH, 1:1000, Millipore, AB1542); Girk2 (1:500, Alomone, APC-006); Calbindin (1:1000, Sigma, C9848), and HuNu (1:1000, Millipore, MaB 1281).

Lelos M.J., Morgan R.J., Kelly C.M., Torres E.M., Rosser A.E, & Dunnett S.B. (2016). Amelioration of non-motor dysfunctions after transplantation of human dopamine neurons in a model of Parkinson's disease. Experimental Neurology, 278, 54-61.

+ Open protocol

+ Expand

Histological Analysis of Aortic Calcification

Check if the same lab product or an alternative is used in the 5 most similar protocols

After organ culture for 7 days, thoracic aorta segments were washed with PBS and fixed with 10% neutral buffer formalin. The specimens were then embedded in paraffin, and 5μm cross-sections were cut. The sections were deparaffinized and H&E staining was performed. Arterial medial calcification was visualized using Alizarin Red S staining as previously described ^{23 (link)}. For immunohistochemistry, the sections were deparaffinized, followed by treatment with citrate buffer for antigen retrieval and 3% H₂O₂. The sections were blocked with Dako serum-free blocking solution (Dako North America, USA) and incubated with primary antibody for overnight at 4°C. The primary antibodies included SM22α (Abcam, ab14106), Osterix (Abcam, ab94744). Subsequently, the sections were incubated with biotinylated secondary antibodies for 30 min at room temperature. Avidin-biotinylated enzyme complex (Vector Laboratories, USA) and a diaminobenzidine substrate chromogen system (Dako North America, USA) were used for detection. Sections were counterstained with hematoxylin. The images were captured by Olympus DP260.

Lin T., Wang X.L., Zettervall S.L., Cai Y, & Guzman R.J. (2016). DMH1, a Highly Selective Small Molecule BMP Inhibitor, Suppresses Arterial Medial Calcification. Journal of vascular surgery, 66(2), 586-593.

+ Open protocol

+ Expand

Immunohistochemical Analysis of B-cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse tissues were fixed in 10% neutral buffered formalin, routinely processed, and embedded in paraffin. Tissues were sectioned at 5 µm and stained with hematoxylin and eosin for histopathological examination. For immunohistochemistry, 5 µm sections were deparaffinized into ethanol and endogenous peroxidase activity was blocked using 0.6% H₂O₂ in methanol. Antigen retrieval was carried out by microwaving in citrate buffer and slides were stained with biotin-conjugated anti-CD45R/B220 (BD Biosciences #553086) 1:200 for B-cell lymphocytes. Detection of CD45R signal was performed using the avidin-biotinylated enzyme complex (Vector Laboratories) with 3,3’-diaminobenzidine (Sigma) as a chromagen. Slides were coun-terstained with hematoxylin. Slides were photographed using an Olympus BX40 microscope, Q-image RGB camera using Bioquant (Nashville, TN) software to capture and achieve the images.

Zhao Z., Fan H., Higgins T., Qi J., Haines D., Trivett A., Oppenheim J.J., Wei H., Li J., Lin H, & Howard O.M. (2014). Fufang Kushen injection inhibits sarcoma growth and tumor-induced hyperalgesia via TRPV1 signaling pathways. Cancer letters, 355(2), 232-241.

+ Open protocol

+ Expand

Immunohistochemical Staining of Cryosectioned Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols

For cryosectioned tissue, 4 µm-thick sections were fixed and incubated with primary antibodies overnight at 4°C, followed by incubation with biotinylated secondary antibodies (Vector Laboratories, USA). Sections were then incubated in Avidin:Biotinylated enzyme Complex (Vector Laboratories) for 30 min, developed with 3,3’-diaminobenzidine substrate (Vector Laboratories) and nuclei counterstained with methyl green. Slides were mounted with Fluka Eukitt® quick-hardening mounting medium (Sigma-Aldrich, USA). See Supplementary Methods for detailed procedure.

He Q., Au B., Kulkarni M., Shen Y., Lim K.J., Maimaiti J., Wong C.K., Luijten M.N., Chong H.C., Lim E.H., Rancati G., Sinha I., Fu Z., Wang X., Connolly J.E, & Crasta K.C. (2018). Chromosomal instability-induced senescence potentiates cell non-autonomous tumourigenic effects. Oncogenesis, 7(8), 62.

+ Open protocol

+ Expand

Quantitative Immunohistochemistry Analysis of IDH, MGMT, and STAT3 in Gliomas

Check if the same lab product or an alternative is used in the 5 most similar protocols

Histology, isocitrate dehydrogenase (IDH) mutation status and O-6-methylguanine-DNA methyltransferase (MGMT) promoter methylation were obtained from the patient's clinical pathology report. Formalin-fixed, paraffin-embedded tumor specimens were transversely sliced into 4 μm sections. Sections were deparaffinized, rehydrated and stained with primary mouse anti-human monoclonal antibodies against p-STAT3 (Cell Signaling Tech, 1:200) and CD3 (Dako, 1:50) overnight. A biotinylated secondary antibody along with avidin/biotinylated enzyme complex (Vector Laboratories) were applied the next day according to the manufacturer's instructions. The color was developed using the ImmPACT DAB enzyme substrate mixture. All sections were counterstained with hematoxylin and eosin (H&E), dehydrated, and mounted. For IHC quantification, the slides were scanned and converted to a digital format with PerkinElmer Automated Quantitative Pathology Imaging System. The low power field (10×) whole slide scan was used for identification of areas of interest in Phenochart and 25% was acquired on high power field (20×). Quantification analysis was performed using inForm Cell Analysis software (ver 2.4). Whole section slides of each marker were reviewed next to the corresponding H&E slide.

de Groot J., Ott M., Wei J., Kassab C., Fang D., Najem H., O'Brien B., Weathers S.P., Matsouka C.K., Majd N.K., Harrison R.A., Fuller G.N., Huse J.T., Long J.P., Sawaya R., Rao G., MacDonald T.J., Priebe W., DeCuypere M, & Heimberger A.B. (2022). A first-in-human Phase I trial of the oral p-STAT3 inhibitor WP1066 in patients with recurrent malignant glioma. CNS Oncology, 11(2), CNS87.

+ Open protocol

+ Expand

Quantifying NF-κB p65 Expression by IHC

Check if the same lab product or an alternative is used in the 5 most similar protocols

One section of each colon was used for immunohistochemistry (IHC) with a specific antibody for NF-κB p65 as described previously^{21 (link)}. In brief, the sections were deparaffinated in xylene and rehydrated, unmasked in antigen unmasking solution (DAKO, Copenhagen, Denmark). Endogenous peroxidase activity was quenched by 3% H₂O₂ in distilled water. Sections were then treated for 1 hour at room temperature in PBS containing 10% normal serum and incubated with primary antibodies: NF-κB p65 (1:2000, Abcam, Cambridge, MA) overnight at 4°C and incubated with biotinylated secondary antibody (1:200 in 10% normal serum). Then, the sections were treated with avidin-biotinylated enzyme complex (Vector Laboratories, Burlingame, CA) and 3,3′-diaminobenzidine (Vector Laboratories). Finally, the sections were counterstained with hematoxylin and mounted with Permount (Sigma-Aldrich, St. Louis, MO). Negative controls were processed in the absence of the primary antibody. The total number of cells and number of positively stained cells were quantified using the Aperio ScanScope GL system (Aperio, Vista, CA). The results were presented as the percentage of the cells with positive stained cells.

Feng S., Dai Z., Liu A., Wang H., Chen J., Luo Z, & Yang C.S. (2017). β-Sitosterol and stigmasterol ameliorate dextran sulfate sodium-induced colitis in mice fed a high fat western-style diet. Food & function, 8(11), 4179-4186.

+ Open protocol

+ Expand

Histopathological Evaluation of Rickettsia Infection

Check if the same lab product or an alternative is used in the 5 most similar protocols

After fixation, tissues were routinely embedded in paraffin, sectioned, and stained with hematoxylin and eosin (H&E) for histopathological evaluation. Tissue sections were evaluated in a randomized, blinded manner by a board-certified veterinary anatomic pathologist. Skin from the inoculation sites and lymph node sections for all groups were assessed by immunohistochemistry (IHC) for the presence of Rickettsia using an anti-RC_PFA polyclonal rabbit primary antibody [41 (link)]. Cross-reactivity of this antibody to R. parkeri was confirmed by staining R. parkeri (Portsmouth strain) infected Vero cells. Briefly, slides were stained using a DAKO autostainer LINK 48 after proteinase K antigen retrieval (Dako, Carpinteria, CA, USA) with anti-RC_PFA (1:2000) and a biotinylated anti-rabbit secondary antibody (Vector Laboratories, Burlingame, CA, USA), and visualized using the avidin/biotinylated enzyme complex (Vector Labs) and the ImmPACT NovaRED peroxidase substrate (Vector Labs), followed by counterstaining with Mayer’s hematoxylin. False positives due to non-specific binding of the secondary antibody were ruled out by comparing sample staining to staining in tissue sections that were stained without primary antibody.

Banajee K.H., Embers M.E., Langohr I.M., Doyle L.A., Hasenkampf N.R, & Macaluso K.R. (2015). Amblyomma maculatum Feeding Augments Rickettsia parkeri Infection in a Rhesus Macaque Model: A Pilot Study. PLoS ONE, 10(8), e0135175.

+ Open protocol

+ Expand

Immunohistochemical Staining of Neuronal Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Free-floating sections were blocked in 3% normal serum, then incubated in primary antibody with 1% serum overnight at room temperature. Tissue was incubated in secondary antibody (1:200) with 1% serum for 3 h, then immersed in an avidin-biotinylated enzyme complex (Vector Laboratories, Peterborough, UK) for 2 h, then stained with either DAB or Vector SG (Vector Labs).
The primary antibodies used were DARPP-32 (human specific, 1:1,000; Abcam, Cambridge, UK); DARPP-32 (1:30,000; kind gift from Cornell University), tyrosine hydroxylase (TH, 1:1,000; Millipore, Watford, UK), b-tubulin (1:500; Sigma-Aldrich, Poole, UK), NeuN (1:1,000; Millipore), and HuNu (1:1,000; Millipore).

Lelos M.J., Roberton V.H., Vinh N.N., Harrison C., Eriksen P., Torres E.M., Clinch S.P., Rosser A.E, & Dunnett S.B. (2016). Direct Comparison of Rat- and Human-Derived Ganglionic Eminence Tissue Grafts on Motor Function. Cell transplantation, 25(4).

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!