Hrp conjugated rat anti ha

Manufactured by Roche

Sourced in United States

The HRP-conjugated rat anti-HA is a laboratory reagent used for the detection and identification of proteins tagged with the HA (hemagglutinin) epitope. It is a conjugate of a rat-derived antibody specific to the HA tag and horseradish peroxidase (HRP), a widely used enzyme label for various immunoassays and detection methods.

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Lab products found in correlation

Q tof ultima api mass spectrometer, by Waters Corporation (1 mentions) 3 3 diaminobenzidine tetrahydrochloride dab, by Merck Group (1 mentions) Sigmafast opd, by Merck Group (1 mentions) Rabbit anti par3, by Merck Group (1 mentions) Methyl β cyclodextrin mβcd, by Merck Group (1 mentions) Hrp conjugated secondary antibody, by GE Healthcare (1 mentions) Chlorpromazine, by LKT Laboratories (1 mentions)

4 protocols using hrp conjugated rat anti ha

Proteomic Analysis of Thyroid Lesions

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Proteins in PTC and goitre tissues were resolved by two-dimensional gel electrophoresis (2D) (Roessler et al, 2006 (link)). Two gels were performed for both the papillary carcinoma and goitre proteins. One was stained with Comassie Brilliant Blue G-250 dye (Bio-Rad, Hercules, CA, USA) and the other was transferred onto a nitrocellulose membrane (GE) for western blot with scFV-C1 antibody, using the HRP-conjugated rat anti-HA (Roche Applied Science). The corresponding spots that scFv antibody recognised in the western blot were excised from the gels and subjected to trypsin digestion (Wilm et al, 1996 (link)), followed by liquid chromatography mass spectrometry (LC-MS/MS) to allow identification.
Peptides were identified by on-line chromatography using a Cap-LC coupled to a Q-TOF Ultima API mass spectrometer (Waters, Milford, MA, USA). The resulting fragmented spectra were processed using the ProteinLynx 4.0 software (Waters), and we used the MASCOT MS/MS Ion Search system to blast the sequences against the SwissProt and NCBInr databases. According to the MASCOT probability method, only significant (P<0.05) hits were accepted.

Carneiro A.P., Reis C.F., Morari E.C., Maia Y.C., Nascimento R., Bonatto J.M., de Souza M.A., Goulart L.R, & Ward L.S. (2014). A putative OTU domain-containing protein 1 deubiquitinating enzyme is differentially expressed in thyroid cancer and identifies less-aggressive tumours. British Journal of Cancer, 111(3), 551-558.

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Immunohistochemistry Analysis of scFv-D09 Affinity in Oral Cancer

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To verify the affinity of scFv-D09 monoclonal antibody in oral squamous carcinoma tissue, immunohistochemistry was performed. For this, paraffin embedded tissue sections were deparaffinized, rehydrated and submitted to heat pretreatment in 0.1 M sodium citrate buffer for 1 h at 90 °C for antigen retrieval. Then, endogenous peroxidase activity was blocked with three 5-min washes with 3% H₂O₂. The sections were rinsed in distilled water and then blocked for 1 h in PBS–10% BSA. Then, approximately 4 µg/µl of the scFv-D09 were incubated in a humidifying chamber for 18 h at 4 °C. After that, slides were washed two times in PBS for 2 min each followed by incubation with HRP conjugated rat anti-HA (Roche Applied Science, Indianapolis, IN, USA) diluted (1:400) for 1 h at room temperature. After a washing step, peroxidase activity was visualized by incubation in 3, 3′- diaminobenzidine tetrahydrochloride (DAB, Sigma-Aldrich, St. Louis, MO, USA) for 5 min at room temperature and counterstained with Harris haematoxylin. Fragments of human lip well differentiated carcinoma were utilized as positive controls, and mucocele as negative control.

Faria P.C., Carneiro A.P., Binato R., Nascimento R., Santos P.S., Fagundes D., da Silva S.J., Loyola A.M., Abdelhay E, & Goulart L.R. (2019). Upregulation of tropomyosin alpha-4 chain in patients’ saliva with oral squamous cell carcinoma as demonstrated by Phage display. Scientific Reports, 9, 18399.

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Screening Thyroid Cancer Antigens by scFv

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Maxisorp 96-well (NUNC) plates were coated with 1.0 μg per well of total pooled protein extracted from goitre, adenoma and tumour thyroid tissues to screen for clones that could differentially recognise thyroid cancer antigens in 100 μl of sodium bicarbonate buffer (pH 7.4) overnight at 4 °C. Wells were washed three times with PBST and then blocked for 1 h at 37 °C with PBS plus 5% nonfat milk (PBSM 5%). After blocking, wells were washed three times with PBST and 4 μg of purified scFvs were added to each well and incubated for 2 h at room temperature under low agitation. Then, the plate was washed three times with PBST, followed by incubation with HRP-conjugated rat anti-HA (Roche Applied Science, Indianapolis, IN, USA) diluted (1 : 1000) in PBSM 5% for 1 h at 37 °C. After another four washes, the chromogenic substrate OPD SigmaFast (Sigma-Aldrich) was added and the absorbance was measured at 492 nm.

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Protein Localization and Inhibition in Podocytes

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The antibodies used in this study were: rabbit anti-nephrin and rabbit anti-podocin (IBL; Immuno-Biological Laboratories, Fujioka, Gunma, Japan), rabbit anti-aPKC and mouse anti-GFP (B-2; Santa Cruz Biotechnology, Dallas, TX, USA), HRP-conjugated rat anti-HA (Roche Diagnostics, Basel, Switzerland), rabbit anti-Par3 (Merck Millipore, Billerica, MA, USA), mouse anti-E-cadherin (36/E-Cadherin) and mouse anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (BD Biosciences, San Jose, CA, USA), mouse anti-transferrin receptor (TfR) (H68.4), mouse anti-zonula occludens-1 (ZO1-1A12), Alexa488 and Alexa555-conjugated secondary antibodies (Life Technologies, Carlsbad, CA, USA), HRP-conjugated secondary antibodies (GE Healthcare Bio-Sciences, Piscataway, NJ, USA) and rabbit anti-neph1 (as previously described (37 (link))). The reagents used in this study were chlorpromazine (LKT Laboratories Inc., St Paul, MN, USA), methyl-β-cyclodextrin (MβCD) (Sigma, St Louis, MO, USA) and doxycycline-HCl (ICN Biochemicals Inc., Irvine, CA, USA). Myristoylated pseudosubstrate inhibitor specific for atypical PKC subtypes (myr-SIYRRGARRWRKL) or scramble peptide (SC) (myr-RLYRKRIWRSAGR) was purchased from Peptide Institute Inc. (Osaka, Japan).

Satoh D., Hirose T., Harita Y., Daimon C., Harada T., Kurihara H., Yamashita A, & Ohno S. (2014). aPKCλ maintains the integrity of the glomerular slit diaphragm through trafficking of nephrin to the cell surface. Journal of Biochemistry, 156(2), 115-128.

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