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Radioimmunoprecipitation Assay

Radioimmunoprecipitation Assay (RIPA) is a powerful analytical technique used to study protein-protein interactions and protein expression levels.
This AI-powered tool from PubCompare.ai helps researchers optimize their RIPA experiments by providing curated comparisons of protocols from literature, preprints, and patents.
The AI-driven analysis identifies the most effective methods and products, empowering scientists to design robust and efficient RIPA experiments.
With PubCompare.ai, researchers can easily locate and assess the best RIPA protocols, streamlining their research and accelerating scientific discoveries.

Most cited protocols related to «Radioimmunoprecipitation Assay»

1
Quantitative Proteomics of HeLa-E. coli Mixture
Cited 284 times
An Escherichia coli K12 strain was grown in standard LB medium, harvested, washed in PBS, and lysed in BugBuster (Novagen Merck Chemicals, Schwalbach, Germany) according to the manufacturer's protocol. HeLa S3 cells were grown in standard RPMI 1640 medium supplemented with glutamine, antibiotics, and 10% FBS. After being washed with PBS, cells were lysed in cold modified RIPA buffer (50 mm Tris-HCl, pH 7.5, 1 mm EDTA, 150 mm NaCl, 1% N-octylglycoside, 0.1% sodium deoxycholate, complete protease inhibitor mixture (Roche)) and incubated for 15 min on ice. Lysates were cleared by centrifugation, and after precipitation with chloroform/methanol, proteins were resuspended in 6 m urea, 2 m thiourea, 10 mm HEPES, pH 8.0. Prior to in-solution digestion, 60-μg protein samples from HeLa S3 lysates were spiked with either 10 μg or 30 μg of E. coli K12 lysates based on protein amount (Bradford assay). Both batches were reduced with dithiothreitol and alkylated with iodoacetamide. Proteins were digested with LysC (Wako Chemicals, GmbH, Neuss, Germany) for 4 h and then trypsin digested overnight (Promega, GmbH, Mannheim, Germany). Digestion was stopped by the addition of 2% trifluroacetic acid. Peptides were separated by isoelectric focusing into 24 fractions on a 3100 OFFGEL Fractionator (Agilent, GmbH, Böblingen, Germany) as described in Ref. 45 (link). Each fraction was purified with C18 StageTips (46 (link)) and analyzed via liquid chromatography combined with electrospray tandem mass spectrometry on an LTQ Orbitrap (Thermo Fisher) with lock mass calibration (47 (link)). All raw files were searched against the human and E. coli complete proteome sequences obtained from UniProt (version from January 2013) and a set of commonly observed contaminants. MS/MS spectra were filtered to contain at most eight peaks per 100 mass unit intervals. The initial MS mass tolerance was 20 ppm, and MS/MS fragment ions could deviate by up to 0.5 Da (48 (link)). For quantification, intensities can be determined alternatively as the full peak volume or as the intensity maximum over the retention time profile, and the latter method was used here as the default. Intensities of different isotopic peaks in an isotope pattern are always summed up for further analysis. MaxQuant offers a choice of the degree of uniqueness required in order for peptides to be included for quantification: “all peptides,” “only unique peptides,” and “unique plus razor peptides” (42 (link)). Here we chose the latter, because it is a good compromise between the two competing interests of using only peptides that undoubtedly belong to a protein and using as many peptide signals as possible. The distribution of peptide ions over fractions and samples is shown in supplemental Fig. S1.
Cox J., Hein M.Y., Luber C.A., Paron I., Nagaraj N, & Mann M. (2014). Accurate Proteome-wide Label-free Quantification by Delayed Normalization and Maximal Peptide Ratio Extraction, Termed MaxLFQ. Molecular & Cellular Proteomics : MCP, 13(9), 2513-2526.
Publication 2014
Acids Antibiotics, Antitubercular Biological Assay Buffers Cells Centrifugation Chloroform Cold Temperature Deoxycholic Acid, Monosodium Salt Digestion Dithiothreitol Edetic Acid Escherichia coli Escherichia coli K12 Glutamine HeLa Cells HEPES Homo sapiens Immune Tolerance Iodoacetamide Ions Isotopes Liquid Chromatography Methanol Peptides Promega Protease Inhibitors Proteins Proteome Radioimmunoprecipitation Assay Retention (Psychology) Sodium Chloride Staphylococcal Protein A Tandem Mass Spectrometry Thiourea Tromethamine Trypsin Urea
2
Evaluating Novel SLE Classification
Cited 247 times
To assess the performance of the new classification rule, we obtained detailed data regarding a new set of 690 additional patients. Sites were again asked to submit information on patients diagnosed with SLE, and on an approximately equal number of controls with the following diagnoses: rheumatoid arthritis, undifferentiated connective tissue disease, primary antiphospholipid antibody syndrome, vasculitis, chronic cutaneous lupus, scleroderma, Sjögren syndrome, myositis, psoriasis, fibromyalgia, alopecia areata and sarcoidosis. These data were collected on standardized case report forms and sent to the coordinating site. Information included a demographics summary, a clinical scenario, specification of ACR criteria that were met and not met, specification of SLICC criteria met and not met, auto-antibody titers and complement titers.
In addition, serum from each patient was sent to the coordinating site and analyzed at the Rheumatology Diagnostic Laboratory (Los Angeles, CA) for anti-dsDNA by ELISA, Crithidia and Farr assays, anti-Smith antibody and complement C3 and C4 levels. A second set of blood samples were tested for antiphospholipid antibodies (lupus anticoagulant, ELISA assay for IgG, IgM and IgA isotypes of anticardiolipin antibodies and anti-beta2 glycoprotein1 antibodies) at the laboratory of Joan Merrill, M.D. (Oklahoma Medical Research Foundation). Direct Coombs was done at each center's own laboratory or at Quest Diagnostics. A short description of each patient (“patient scenario”) was generated containing the submitted information and the updated auto-antibody and complement profiles.
Petri M., Orbai A.M., Alarcón G.S., Gordon C., Merrill J.T., Fortin P.R., Bruce I.N., Isenberg D., Wallace D.J., Nived O., Sturfelt G., Ramsey-Goldman R., Bae S.C., Hanly J.G., Sanchez-Guerrero J., Clarke A., Aranow C., Manzi S., Urowitz M., Gladman D., Kalunian K., Costner M., Werth V.P., Zoma A., Bernatsky S., Ruiz-Irastorza G., Khamashta M.A., Jacobsen S., Buyon J.P., Maddison P., Dooley M.A., van Vollenhoven R.F., Ginzler E., Stoll T., Peschken C., Jorizzo J.L., Callen J.P., Lim S.S., Fessler B.J., Inanc M., Kamen D.L., Rahman A., Steinsson K., Franks AG J.r., Sigler L., Hameed S., Fang H., Pham N., Brey R., Weisman M.H., McGwin G J.r, & Magder L.S. (2012). Derivation and Validation of Systemic Lupus International Collaborating Clinics Classification Criteria for Systemic Lupus Erythematosus. Arthritis and rheumatism, 64(8), 2677-2686.
Publication 2012
Alopecia Anti-Antibodies Antibodies, Anti-Idiotypic Antiphospholipid Antibodies Antiphospholipid Syndrome Biological Assay BLOOD Complement 3 Crithidia Dermatosclerosis Diagnosis Direct Coombs Test DNA, Double-Stranded Enzyme-Linked Immunosorbent Assay Fibromyalgia Immunoglobulin Isotypes Immunoglobulins Lupus Coagulation Inhibitor Lupus Erythematosus, Chronic Cutaneous Myositis Patients Psoriasis Radioimmunoprecipitation Assay Rheumatoid Arthritis Sarcoidosis Serum Sjogren's Syndrome Undifferentiated Connective Tissue Diseases Vasculitis
3
Quantifying Protein-Protein Interactions
Cited 55 times

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Publication 2012
Buffers Capillaries Cells HEK293 Cells Magnesium Chloride Proteins Radioimmunoprecipitation Assay Sodium Chloride Technique, Dilution Tromethamine Tween 20
4
ChIPmentation: Versatile Chromatin Profiling
Cited 51 times
ChIPmentation is compatible with various different protocols for ChIP, which makes it easy to apply ChIP-mentation to antibodies that work best with a certain ChIP protocol. The chosen ChIP protocol was followed until the beads carrying immunoprecipitated chromatin were washed with LiCl-containing wash buffer (WBIII for ChIP version 1, RIPA-LiCl for ChIP version 2, and TF-WBIII for ChIP version 3, as described in the Supplementary Note). Beads were then washed twice with 10 mM cold Tris-Cl pH 8.0 to remove detergent, salts, and EDTA. Subsequently, beads were resuspended in 30 μl of the tagmentation reaction buffer (10 mM Tris, pH 8.0, 5 mM MgCl2) containing 1 μl Tagment DNA Enzyme from the Nextera DNA Sample Prep Kit (Illumina) and incubated at 37°C for 10 minutes in a thermocycler. Following tagmentation, the beads were washed twice with 150 μl cold WBI (ChIP version 1), RIPA (ChIP version 2), or TF-WBI (ChIP version 3). Afterward, the chosen ChIP protocol was resumed with the final bead wash, elution from beads, reverse-crosslinking, and DNA purification. A detailed protocol can be found at http://chipmentation.computational-epigenetics.org/.
Schmidl C., Rendeiro A.F., Sheffield N.C, & Bock C. (2015). ChIPmentation: fast, robust, low-input ChIP-seq for histones and transcription factors. Nature methods, 12(10), 963-965.
Publication 2015
Antibodies Buffers Chromatin Cold Temperature Detergents DNA Chips Edetic Acid Enzymes Epigenetic Process Magnesium Chloride Radioimmunoprecipitation Assay Salts Tromethamine
5
Western Blot Quantification of Cellular Signaling Proteins
Cited 45 times
The MCF-7 cell line was maintained under standard conditions in Dulbecco's modified Eagle's medium supplemented with 10% foetal bovine serum. Cells were washed with ice cold phosphate buffered Saline and lysed in RIPA buffer (1% NP-40, 0.1% SDS, 0.5% Sodium deoxycholate, 50 mM Tris pH 7.5, 150 mM NaCl) supplemented with protease and phosphatase inhibitor cocktails (Sigma Aldrich) and protein concentration was quantitated by BCA protein assay (Invitrogen). Purified BSA (Applichem) was dissolved in RIPA buffer. Cell lysates and a BSA sample were serially diluted 1∶2 and run on SDS-PAGE using a standard protocol. Proteins were transferred to the PVDF (for ECL based detection) or Nitrocellulose (for LI-COR based proteins detection) membranes. Membranes were blocked with blocking solution (11500694001, Roche) for BSA detection or 5% skimmed milk for rest of the membranes. For Western blotting ERK (M-5670, Sigma Aldrich), mTOR (2972, Cell Signaling Technology), RSK1 (sc-231, Santa Cruz) and BSA (sc-50528, Santa Cruz) antibodies were used. Anti-rabbit HRP-conjugated (Cell Signaling Technology) or anti-Rabbit IR 800 (LI-COR) secondary antibodies were used for ECL or LI-COR protein detection systems, respectively. Signal was detected by standard X-ray films (Fuji), CCD camera (Advanced Molecular Vision) or LI-COR scanner.
Degasperi A., Birtwistle M.R., Volinsky N., Rauch J., Kolch W, & Kholodenko B.N. (2014). Evaluating Strategies to Normalise Biological Replicates of Western Blot Data. PLoS ONE, 9(1), e87293.
Publication 2014
Antibodies Biological Assay Buffers Cells Cold Temperature Deoxycholic Acid, Monosodium Salt Fetal Bovine Serum FRAP1 protein, human MCF-7 Cells Milk, Cow's Nitrocellulose Nonidet P-40 Peptide Hydrolases Phosphates Phosphoric Monoester Hydrolases polyvinylidene fluoride Proteins Rabbits Radioimmunoprecipitation Assay RPS6KA1 protein, human Saline Solution SDS-PAGE Sodium Chloride Tissue, Membrane Tromethamine Vision X-Ray Film

Most recents protocols related to «Radioimmunoprecipitation Assay»

1
EV Protein Isolation and Western Blot
Total protein was isolated from EVs using Pierce RIPA lysis buffer with Halt protease and phosphatase inhibitor cocktail (Thermo Fisher, Waltham, MA), and protein concentration was determined using the Pierce BCA protein assay kit (Thermo Fisher). For Western blotting, samples were prepared in Laemmli buffer, boiled at 95 °C for 5 min, and 5 μg protein was loaded. Proteins were separated by SDS-PAGE using 4–15% TGX Gels (Criterion, Bio-Rad, Hercules, CA) by running at 200 V at room temperature. Proteins were transferred for 60 min at 100 V on ice onto a nitrocellulose membrane in 20% methanol Tris-glycine buffer. The Revert Total Protein Stain Kit (Li-Cor Biosciences, Lincoln, NE) or Ponceau S solution (Thermo Fisher) was used to stain total protein, and the membranes were imaged to verify transfer efficiency and loading. The membranes were subsequently blocked in 5% nonfat dry milk in Tris-buffered saline-Tween (TBS-T, 0.1% Tween-20) for 1 h at room temperature, then incubated overnight at 4 °C in primary antibody (anti-CD63, EXOAB-CD63A-1; System Biosciences, Palo Alto, CA and anti-Apolipoprotein A1 (ApoA1, 701239; Thermo Fisher) at a 1:1000 dilution in 5% nonfat dry milk in TBS-T. The membranes were then washed before incubation in goat anti-rabbit secondary antibodies (EXOAB-CD63A-1; System Biosciences) (1:10,000 dilution) for 1 h at room temperature. Blots were developed with enhanced chemiluminescence (Clarity Western ECL Substrate, Bio-Rad), imaged, and quantified with ImageJ (National Institutes of Health).
Ismaeel A., Van Pelt D.W., Hettinger Z.R., Fu X., Richards C.I., Butterfield T.A., Petrocelli J.J., Vechetti I.J., Confides A.L., Drummond M.J, & Dupont-Versteegden E.E. (2023). Extracellular vesicle distribution and localization in skeletal muscle at rest and following disuse atrophy. Skeletal Muscle, 13, 6.
Publication 2023
Anti-Antibodies APOA1 protein, human Biological Assay Buffers Chemiluminescence Gels Glycine Goat Immunoglobulins Laemmli buffer Methanol Milk, Cow's Nitrocellulose Peptide Hydrolases Phosphoric Monoester Hydrolases ponceau S Proteins Rabbits Radioimmunoprecipitation Assay Saline Solution SDS-PAGE Stains Technique, Dilution Tissue, Membrane Tween 20 Tweens
2
Western Blot Analysis of Inflammatory and Tight Junction Proteins
Proteins were extracted from colon tissues with RIPA lysis buffer (Beyotime, Shanghai, China) containing phenylmethyl sulfonyl fluoride. A bicinchoninic acid protein assay kit (Beyotime) was used to measure the concentration of protein. Suitable quality protein samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to PVDF membranes (Millipore, Billerica, MA, USA). The PVDF membranes were blocked with 8% skim milk at room temperature for 1 h and incubated with primary antibodies at 4 °C overnight. The corresponding secondary antibodies were used at room temperature for 1 h, and the protein signals were detected with a FluorChem FC3 system (ProteinSimple, California, USA) using an enhanced chemilusystem reagent (Thermo Fisher, Waltham, USA) according to the manufacturer’s instructions. The antibodies used in this study were as follows: TNFα (1:200, Santa Cruz Biotechnology, Texas, USA), IL-1β (1:1000, Cell Signaling Technology, Massachusetts, USA), IL6 (1:200, Santa Cruz Biotechnology), ZO-1 (1:1000, Invitrogen, California, USA), Occludin (1:1000, Invitrogen), P62 (1:10,000, Abcam, Cambridge, USA), LC3 (1:1000, Proteintech, Chicago, America), CB1 (1:1000, Proteintech), GAPDH (Abclonal, Wuhan, China), horseradish peroxidase (HRP)-linked goat anti-rabbit IgG, and HRP-linked goat anti-mouse IgG (1:4000, Antgene, Wuhan, China).
Yang J., Wang L., Mei M., Guo J., Yang X, & Liu S. (2023). Electroacupuncture repairs intestinal barrier by upregulating CB1 through gut microbiota in DSS-induced acute colitis. Chinese Medicine, 18, 24.
Publication 2023
anti-IgG Antibodies bicinchoninic acid Biological Assay Buffers Colon GAPDH protein, human Goat Horseradish Peroxidase Interleukin-1 beta Milk, Cow's Mus Occludin polyvinylidene fluoride Proteins Rabbits Radioimmunoprecipitation Assay SDS-PAGE sulfuryl fluoride Tissue, Membrane Tissues Tumor Necrosis Factor-alpha
3
Western Blot Analysis of Apoptosis and MAPK Signaling
Total proteins from BC cells were extracted with radioimmunoprecipitation assay (RIPA) lysis buffer containing phenylmethyl sulfonylfluoride (PMSF), followed by SDS-PAGE, and transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Billerica, MA, USA). After that, the PVDF membranes were blocked with 5% skimmed milk and incubated with the corresponding primary antibody: Bax (1:1000, Abcam, Cambridge, UK), Bcl-2 (1:1000, Abcam), MAPK6 (1:1000, Abcam), p-MAPK6 (1:1000, Abcam), p38 (1:1000, Cell Signaling Technology, Danvers, MA, USA), p-p38 (1:1000, Cell Signaling Technology), ERK (1:1000, Cell Signaling Technology), p-ERK (1:1000, Cell Signaling Technology), and β-actin (1:5000, Cell Signaling Technology). Next, the membranes were incubated at 4℃ for 14 h, and subsequently, the membranes were incubated with the corresponding secondary antibody (1:6000, Cell Signaling Technology) for 1.5 h at room temperature. Lastly, the band signal was visualized using an enhanced chemiluminescence (ECL) detection system (Millipore).
Wang B., Chen H., Deng Y., Chen H., Xing L., Guo Y., Wang M, & Chen J. (2023). CircDNAJC11 interacts with TAF15 to promote breast cancer progression via enhancing MAPK6 expression and activating the MAPK signaling pathway. Journal of Translational Medicine, 21, 186.
Publication 2023
Actins BCL2 protein, human Buffers Cells Chemiluminescence Immunoglobulins Milk, Cow's Mitogen-Activated Protein Kinase 3 polyvinylidene fluoride Proteins Radioimmunoprecipitation Assay SDS-PAGE sulfuryl fluoride Tissue, Membrane
4
Protein Expression Analysis by Western Blot
Total protein was isolated from RIPA lysate and quantified using the BCA method. Next, 30 μg of total protein was separated by 8% sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred onto a polyvinylidene fluoride membrane. After blocking with 5% skim milk at 37 °C for 2 h, the membrane was incubated with primary antibodies against LDHA (1:1 000), CD47 (1:1 000), PDIA3 (1:1 000), SLC16A1 (1: 200), and the internal reference β-actin (1:1 000) overnight at 4 °C. The next day, the membrane was incubated with secondary antibodies at 37 °C for 1 h. The bands were detected by adding the chemiluminescence substrate, and proteins were quantified by densitometry using Image Lab 5.2 software, with β-actin as the internal reference. The experiment was performed three times.
Zhang H., Wang X., Li Y., Bai Y., Li Q., Wang S., Wei Y., Li J., Wen S, & Zhao W. (2023). The hsa_circ_0000276-ceRNA regulatory network and immune infiltration in cervical cancer. BMC Cancer, 23, 222.
Publication 2023
Actins Antibodies CD47 protein, human Chemiluminescence Densitometry LDH 5 Milk, Cow's PDIA3 protein, human polyvinylidene fluoride Proteins Radioimmunoprecipitation Assay SDS-PAGE Tissue, Membrane
5
Pancreatic Cancer Cell Culture Protocols
Pancreatic cancer cell lines (AsPC-1 and BxPC-3) were cultured in RPMI-1640 (Corning, NY, USA) with 10% fetal bovine serum (FBS) and 1% penicillin–streptomycin. Two additional pancreatic cancer cell lines (PANC-1, MIA Paca-2) were cultured in DMEM (Dulbecco’ modified eagle medium) (Gibco, Grand Island, NY, USA) supplemented with 10% FBS and 1% penicillin–streptomycin. Human pancreatic ductal epithelium (hTERT-HPNE) cells were cultured in Medium D with mixtures of M3 and DMEM medium containing one volume of medium M3TM Base F culture media (InCell Corp., San Antonio, TX, USA), three volumes of glucose-free DMEM, 5% FBS, 5.5 mM glucose, 10 ng/ml EGF, and 50 µg/ml gentamycin [26 (link)]. All these cells were cultured at 37 °C in a humidified atmosphere containing 5% CO2. RNA was extracted from tissues using the TRIzol reagent (Invitrogen, Carlsbad, CA, USA) and was reverse-transcribed into cDNA using the PrimeScript RT Master Mix (Takara, Otsu, Shiga, Japan). RT-qPCR analyses were quantified with PowerUp™ SYBR® Green Master Mix (Applied Biosystems, Austin, TX, USA), and expression levels were normalized to GAPDH levels. Proteins were extracted in RIPA buffer supplemented with a complete, EDTA-free protease and phosphatase inhibitor single-use cocktail (Thermo Scientific). Proteins were separated by SDS-PAGE and blotted onto a PVDF membrane. Anti-TSC22D2 (1:1000 dilution, #25,418–1-AP, Proteintech) was used as primary antibodies for immunoblotting. Reacted antibodies were detected using an enhanced chemiluminescence detection system.
Liu Q., Li R., Wu H, & Liang Z. (2023). A novel cuproptosis-related gene model predicts outcomes and treatment responses in pancreatic adenocarcinoma. BMC Cancer, 23, 226.
Publication 2023
Antibodies Atmosphere austin Buffers Cell Lines Cells Chemiluminescence Culture Media DNA, Complementary Eagle Edetic Acid Epithelium Fetal Bovine Serum GAPDH protein, human Gentamicin Glucose Homo sapiens Pancreatic Cancer Pancreatic Duct Penicillins Peptide Hydrolases Phosphoric Monoester Hydrolases polyvinylidene fluoride Proteins Radioimmunoprecipitation Assay SDS-PAGE Streptomycin SYBR Green I Technique, Dilution Tissue, Membrane Tissues trizol

Top products related to «Radioimmunoprecipitation Assay»

1
Pvdf membrane by Merck Group
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PVDF membranes are a type of laboratory equipment used for a variety of applications. They are made from polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF membranes are known for their high mechanical strength, thermal stability, and resistance to a wide range of chemicals. They are commonly used in various filtration, separation, and analysis processes in scientific and research settings.
2
Ripa lysis buffer by Beyotime
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RIPA lysis buffer is a detergent-based buffer solution designed for the extraction and solubilization of proteins from cells and tissues. It contains a mixture of ionic and non-ionic detergents that disrupt cell membranes and solubilize cellular proteins. The buffer also includes additional components that help to maintain the stability and activity of the extracted proteins.
3
Ripa buffer by Beyotime
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RIPA buffer is a detergent-based cell lysis and extraction reagent. It is used to extract and solubilize proteins from cells and tissues for analysis. The buffer contains a combination of ionic and non-ionic detergents, as well as other components that aid in the solubilization and stabilization of proteins.
4
Ripa buffer by Merck Group
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RIPA buffer is a widely used lysis buffer for extracting proteins from cells and tissues. It is a detergent-based buffer that helps solubilize proteins, disrupt cell membranes, and maintain the integrity of protein structures during the extraction process. The buffer contains a combination of ionic and non-ionic detergents, as well as other components that help stabilize and preserve the extracted proteins.
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RIPA buffer is a commonly used lysis buffer for the extraction and solubilization of proteins from cells and tissues. It contains a mixture of ionic and non-ionic detergents that help disrupt cell membranes and release cellular contents, including proteins. The buffer also includes salts and buffers to maintain the pH and ionic strength of the solution.
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Protease inhibitor cocktail by Roche
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Protease inhibitor cocktail is a laboratory reagent used to inhibit the activity of proteases, which are enzymes that break down proteins. It is commonly used in protein extraction and purification procedures to prevent protein degradation.
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Bca protein assay kit by Beyotime
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The BCA protein assay kit is a colorimetric-based method for the quantitative determination of total protein concentration in a sample. It uses bicinchoninic acid (BCA) to detect and quantify the presence of protein.
8
Bca protein assay kit by Thermo Fisher Scientific
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The BCA Protein Assay Kit is a colorimetric detection and quantification method for total protein concentration. It utilizes bicinchoninic acid (BCA) for the colorimetric detection and quantification of total protein. The assay is based on the reduction of Cu2+ to Cu1+ by protein in an alkaline medium, with the chelation of BCA with the Cu1+ ion resulting in a purple-colored reaction product that exhibits a strong absorbance at 562 nm, which is proportional to the amount of protein present in the sample.
9
Protease inhibitor cocktail by Merck Group
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The Protease Inhibitor Cocktail is a laboratory product designed to inhibit the activity of proteases, which are enzymes that can degrade proteins. It is a combination of various chemical compounds that work to prevent the breakdown of proteins in biological samples, allowing for more accurate analysis and preservation of protein integrity.
10
Pierce bca protein assay kit by Thermo Fisher Scientific
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The Pierce BCA Protein Assay Kit is a colorimetric-based method for the quantification of total protein in a sample. It utilizes the bicinchoninic acid (BCA) reaction, where proteins reduce Cu2+ to Cu+ in an alkaline environment, and the resulting purple-colored reaction is measured spectrophotometrically.

More about "Radioimmunoprecipitation Assay"

Radioimmunoprecipitation Assay (RIPA) is a widely used analytical technique in molecular biology and biochemistry.
It is a powerful tool for studying protein-protein interactions and quantifying protein expression levels.
RIPA is often employed in conjunction with other techniques, such as Western blotting, to provide a comprehensive understanding of protein function and regulation.
The RIPA process involves the use of a detergent-based lysis buffer, commonly referred to as RIPA buffer, which helps to solubilize and extract proteins from cells or tissues.
The RIPA buffer typically contains a combination of ionic and non-ionic detergents, as well as other components like Tris-HCl, NaCl, EDTA, and sodium deoxycholate.
These ingredients help to disrupt cell membranes and release the proteins of interest.
Once the proteins are extracted, they are incubated with specific antibodies that recognize the target proteins.
The antibody-protein complexes are then captured using beads coated with Protein A or Protein G, which have a high affinity for the Fc region of the antibodies.
This process is known as immunoprecipitation.
The immunoprecipitated proteins can then be analyzed using various techniques, such as Western blotting, mass spectrometry, or enzymatic assays.
RIPA is particularly useful for studying post-translational modifications, protein interactions, and protein expression changes in response to different stimuli or conditions.
To optimize RIPA experiments, researchers can utilize tools like PubCompare.ai, which leverages artificial intelligence to provide curated comparisons of RIPA protocols from the literature, preprints, and patents.
This AI-powered resource helps scientists identify the most effective methods and products, empowering them to design robust and efficient RIPA experiments.
Additionally, researchers may use complementary techniques, such as PVDF membranes for Western blotting, BCA protein assay kits for quantifying protein concentrations, and protease inhibitor cocktails to prevent protein degradation during the RIPA process.
By combining these tools and techniques, scientists can gain a deeper understanding of the complex protein networks and pathways within cells.