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Antibody Specificity

Antibody Specificity: The degree to which an antibody recognizes and binds to a particular antigen.
Optimizing antibody specificity is crucial for effective research, diagnosis, and treatment.
PubCompare.ai offers AI-powered tools to streamline this process, enabling researchers to discover the best protocols and products by analyzing literature, pre-prints, and patents.
Enhance your antibody specificity research with PubCompare.ai's insights and streamline your workflow for improved results.

Most cited protocols related to «Antibody Specificity»

Chromatin Immunoprecipitation (ChIP) Protocol

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Publication 2009

Antibodies Antibody Specificity cDNA Library Cells Chromatin Chromatin Immunoprecipitation Sequencing DNA, A-Form DNA Chips Formaldehyde Histone Code HSP40 Heat-Shock Proteins Immune Sera Immunoglobulins Proteins Real-Time Polymerase Chain Reaction Tissues Transients Vertebrates

Solexa DNA Sequencing Protocol

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Publication 2008

2',5'-oligoadenylate Antibodies Antibody Specificity Cells DNA Chips Electrophoresis, Agar Gel Genome Medical Devices Mice, House

Plasma p-Tau231 Biomarker Assay Development

All plasma p-tau biomarkers in this study were measured using in-house Single molecule array (Simoa) methods on the HD-X instrument (Quanterix) at the Clinical Neurochemistry Laboratory, Sahlgrenska University Hospital, Mölndal, Sweden. Plasma p-tau181 and plasma NfL measurements used in this study have been previously reported [9 (link), 29 (link)].
For the novel plasma p-tau231 Simoa assay, monoclonal mouse antibodies were generated using a synthetic peptide (K₂₂₄KVAVVR(pT)PPKSPSSAK₂₄₀C) as a KLH-coupled antigen, numbered according to full-length tau-441 phosphorylated on threonine 231. Candidate hybridomas were selected on brain extracts of AD and control brain tissue. The final cloned and purified monoclonal antibody, ADx253, was characterized on synthetic peptides spanning amino acids threonine 217 till serine 241 of full-length tau for its affinity, its phospho-specificity using both phosphorylated and non-phosphorylated peptides and its preferred selectivity in which position 232 was replaced by a Pip, to simulate cis-selectivity of ADx253. A biotin-conjugated N-terminal anti-tau mouse monoclonal antibody was used for detection. Full-length recombinant tau 441 phosphorylated in vitro by glycogen synthase kinase 3β was used as the calibrator. The assay validation focused on dilution linearity, spike recovery, antibody specificity, precision and lower limit of quantification (LLOQ) are described in the supplementary methods, supplementary Table 1–3 and supplementary Fig. 1–2, online resource.

Ashton N.J., Pascoal T.A., Karikari T.K., Benedet A.L., Lantero-Rodriguez J., Brinkmalm G., Snellman A., Schöll M., Troakes C., Hye A., Gauthier S., Vanmechelen E., Zetterberg H., Rosa-Neto P, & Blennow K. (2021). Plasma p-tau231: a new biomarker for incipient Alzheimer’s disease pathology. Acta Neuropathologica, 141(5), 709-724.

Publication 2021

Amino Acids Antibody Specificity Biological Assay Biological Markers Biotin Brain Clinical Laboratory Services Genetic Selection Glycogen Synthase Kinase 3 beta Hybridomas keyhole-limpet hemocyanin Mice, House Monoclonal Antibodies Peptides Plasma Serine Technique, Dilution Threonine Tissues

Validating Specificity of Synaptic Antibodies

The specificity of nc82 against BRP protein has been demonstrated by: 1) the expression pattern of GFP-tagged bruchpilot driven under tissue-specific drivers, which matches nc82 signals in wing discs and tracheal cells, and is also targeted to the active zone of larval NMJ boutons (Wagh et al., 2006 (link)); 2) western blots of adult head extracts using nc82 (Wagh et al., 2006 (link)); and 3) the loss of immune-expression in brp mutant neuromuscular junctions and rescue by expression of BRP in brp mutants (Kittel et al., 2006 (link)). nc82 has been widely used to label synaptic sites in Drosophila, based largely on the pattern of labeling demonstrated at neuromuscular junctions and fly photoreceptor synapses (Hamanaka and Meinertzhagen, 2010 (link)), but reports of its specificity are mostly not complete for synapses of the CNS. In particular, nc82 labels the platform of the T-bar ribbon, and not only has nc82 not been shown to label the platforms at CNS synapses, but not all synapses in the CNS have such organelles (Butcher et al., 2012 (link)).
The specificities of the three epitope-tagged antibodies, rat anti-FLAG, rabbit anti-hemagglutinin (anti-HA), and mouse anti-V5, are validated by the internal controls of the flip-out approach in that: 1) expression patterns differ by GAL4 line and 2) the extent of labeling varies from no label to dense label even though the GAL4 drivers are reasonably broad.

Wolff T., Iyer N.A, & Rubin G.M. (2014). Neuroarchitecture and neuroanatomy of the Drosophila central complex: A GAL4-based dissection of protocerebral bridge neurons and circuits. The Journal of Comparative Neurology, 523(7), 997-1037.

Publication 2014

Adult Antibody Specificity Cells Drosophila Epitopes Head Hemagglutinin Larva Mus Neuromuscular Junction Organelles Photoreceptor Cells Presynaptic Terminals Proteins Rabbits Synapses Tissue Specificity Trachea Western Blot

Immunoblot and Immunohistochemical Analysis of Keratins

For immunoblot analyses, known quantities of recombinant human keratins were electrophoresed, transferred to nitrocellulose, and the blots were incubated with primary antisera diluted in blocking buffer (Tris-buffered saline with 0.5% Tween 20 and 5% powdered milk). Bound primary antibodies were revealed by alkaline phosphatase–conjugated secondary antibodies as recommended by the manufacturer (Bio-Rad Laboratories). The primary antisera for K17 and K6 were prepared as described below. Immunohistochemical analyses were performed on 5-μm sections prepared from either paraffin-embedded or fresh-frozen tissues. The sections were first reacted with the primary antisera and then revealed with either a peroxidase-based reaction (Kirkegaard and Perry Laboratories, Gaithersburg, MD) or by indirect immunofluorescence (Jackson Immunological Reagents, West Grove, PA). In addition to the antisera mentioned above, other primary antisera used in these studies were: a guinea pig anti-K8/18 purchased from ARP laboratories (Belmont, MA); a mouse monoclonal anti-K10 (clone K8.60) from Sigma Chemical Co.; a mouse monoclonal anti-K14 (LL001) donated by I. Leigh (London Hospital Medical College, London, UK) (Purkis et al., 1990 (link)); and a rabbit anti–lef-1 donated by R. Grosschedl (van Genderen et al., 1994 (link)). Double-immunofluorescence staining using primary antibodies from the same species (rabbit anti-K17 and rabbit anti–lef-1) was performed as described (Lewis-Carl et al., 1993 (link)).

McGowan K.M, & Coulombe P.A. (1998). Onset of Keratin 17 Expression Coincides with the Definition of Major Epithelial Lineages during Skin Development. The Journal of Cell Biology, 143(2), 469-486.

Publication 1998

Alkaline Phosphatase Antibodies Antibody Specificity Buffers Cavia porcellus Clone Cells Cytokeratin Fluorescent Antibody Technique Freezing Homo sapiens Immune Sera Immunoblotting Indirect Immunofluorescence LEF1 protein, human Mice, House Milk, Cow's Nitrocellulose Paraffin Embedding Peroxidase Rabbits Saline Solution Tissues Tween 20

Most recents protocols related to «Antibody Specificity»

Quantifying Glycated CD59 Peptide

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Example 4

A sandwich-type ELISA assay is prepared that specifically quantifies GCD59 relative to the glycated CD59 (GCD59) peptide hybrid surrogate. Antibody H9 is used as a capture antibody to capture CD59 in samples tested. The detection antibody is developed using a peptide antigen containing a glucitollysine residue in the equivalent position of K41 in CD59. For this reason, these assays detect the reduced form of GCD59. These anti-glucitollysine GCD59 ELISA kit assays also include a sample preparation step that utilizes NaBH₄as the reducing agent for transformation of the glycated form of GCD59 into the reduced glycated form of GCD59. Kits include solutions of reducing agent in organic solvent to carry out this step.

Sandwich-type Amadori-modified GCD59 ELISA assays do not need sample preparation by NaBH₄reduction due to detection antibody specificity for the non-reduced Amadori-modified form of GCD59. Sample treatment with DTT may, however, improve antibody affinity.

US11866506B2. Anti-CD59 antibodies (2024-01-09). Mellitus, LLC [US]. Inventors: Michael Chorev [US], Jose A. Halperin [US].

Patent 2024

Antibody Affinity Antibody Specificity Antigens Biological Assay CD59 protein, human Enzyme-Linked Immunosorbent Assay glucitollysine Hybrids Immunoglobulins Peptides Reducing Agents Solvents Specimen Handling

Generation of Anti-IL-4R Antibodies

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Example 3

Human anti-IL-4R antibodies were generated as described in U.S. Pat. No. 7,608,693. The exemplary IL-4R antibody used in the following example is a mouse antibody specific for mouse IL-4R and has the following amino acid sequences: a heavy chain variable region (HCVR) comprising SEQ ID NO: 335 and a light chain variable domain (LCVR) comprising SEQ ID NO: 336. The human anti-IL-4R antibody, referred to as dupilumab, specifically binds to human IL-4Rα and comprises a heavy chain variable region (HCVR) comprising SEQ ID NO: 337 and a light chain variable region (LCVR) comprising SEQ ID NO: 338, a heavy chain complementarity determining region 1 (HCDR1) comprising SEQ ID NO: 339, a HCDR2 comprising SEQ ID NO: 340, a HCDR3 comprising SEQ ID NO: 341, a light chain complementarity determining region 1 (LCDR1) comprising SEQ ID NO: 342, a LCDR2 comprising SEQ ID NO: 343 and a LCDR3 comprising SEQ ID NO: 344. The full-length heavy chain of dupilumab is shown as SEQ ID NO: 345 and the full length light chain is shown as SEQ ID NO: 346.

US11866503B2. Methods for treating inflammatory conditions of the airway or lungs by administering antagonist monoclonal antibodies to interleukin-33 and interleukin-4 receptor (2024-01-09). Regeneron Pharmaceuticals, Inc. [US]. Inventors: Jamie M. Orengo [US], Jeanne Allinne [FR], Andrew J. Murphy [US], George D. Yancopoulos [US].

Patent 2024

Amino Acid Sequence antagonists Antibodies Antibodies, Anti-Idiotypic Antibody Specificity Complementarity Determining Region 1 dupilumab Homo sapiens Immunoglobulins Light Mus

Specificity of Anti-Amadori hCD59 Antibodies

Not available on PMC !

Example 3

The specificity of anti-Amadori-modified hGCD59 mAb D2 and D3 to glycated hCD59 was tested in cell lysates derived from diabetic transgenic mice expressing hCD59 transgene (Tg) in red blood corpuscles (RBCs). Diabetes was induced in the hCD59 transgenic mice by administering one dose of Streptozotein (STZ) and blood glucose was measured after two weeks. A mouse was considered diabetic if its blood sugar level was greater than 200 mg/dL. RBCs were obtained from diabetic transgenic mice (D) and control non-diabetic mice (ND), lysed and proteins were extracted. The protein samples were separated using SDS-PAGE (Sodium Dodecyl sulfate-Polyacrylamide gel electrophoresis) and immunoblotted with anti-Amadori modified hGCD59 antibody (D2) and anti hCD59 antibody (FIG. 2). Consistent with the elevation of glycated CD59 in diabetes, the anti-Amadori modified hGCD59 antibody showed an intense band in the diabetic mice and only a faint band in the control non-diabetic mice. Both the diabetic and control non-diabetic mice showed similar levels of hCD59, demonstrating the specificity of the anti-Amadori modified hGCD59 antibody.

US11866506B2. Anti-CD59 antibodies (2024-01-09). Mellitus, LLC [US]. Inventors: Michael Chorev [US], Jose A. Halperin [US].

Patent 2024

Antibodies, Anti-Idiotypic Antibody Specificity Blood Glucose CD59 protein, human Cells Diabetes Mellitus Erythrocytes Mice, Laboratory Mice, Transgenic Proteins SDS-PAGE Syncope Transgenes Western Blotting

Reducing Experimental Bias in Immunoblot Assays

To reduce experimental bias and enhance rigor, all data were analyzed by a genotype-, case-, or treatment-blinded observer. A peptide blocking test was used to confirm pTDP-43 antibody specificity and to prevent analysis of non-target antigenic bands on immunoblots (pTDP-43 blocking peptide: #22309-1-BP, Proteintech). Other antibodies were validated using “orthogonal” or “independent antibody validation” tests (Uhlen et al., 2016 (link)). Specificity of CK2 activity assays was verified using a purified CK2 enzyme (#CY-E1170-1, Cyclex) and a CK2-specific antagonist (CX-4945).

Ohene-Nyako M., Nass S.R., Richard H.T., Lukande R., Nicol M.R., McRae M., Knapp P.E, & Hauser K.F. (2023). Casein Kinase 2 Mediates HIV- and Opioid-Induced Pathologic Phosphorylation of TAR DNA Binding Protein 43 in the Basal Ganglia. ASN NEURO, 15, 17590914231158218.

Publication 2023

Antibodies Antibody Specificity Antigens Biological Assay Cardiac Arrest CX 4945 Enzymes Genotype Immunoblotting Immunoglobulins Muscle Rigidity

Antibody Validation for Histone Modifications

Antibody against anti-histone H2A.Z (1:5000; ab4626) was purchased from Abcam; antibody against Rpd3 (1:500; sc-514160) was purchased from Santa Cruz Biotechnology; antibodies against GFP (1:5000; 66002-1-1g), Myc (1:5000; 60003-2-1g), goat polyclonal anti-mouse IgG (1:5000; SA00001-1), and goat polyclonal anti-rabbit IgG (1:5000; SA00001-2) were obtained from Proteintech; antibody against pan–acetyl-lysine (1:2000, 9441S) was purchased from Cell Signaling Technology; antibody against FLAG M2 (1:3000; F1804-1MG) was obtained from Sigma-Aldrich; antibody against CBP (1:2000; Abs130593) was purchased from Absin Bioscience Inc.; and antibodies against Ino80 (1:500) and Ino80 K929ac (1:500) were custom-made in Abclonal. The specificity of the custom-made antibodies was confirmed by immunoblot analysis with cell extract or IP of corresponding mutants (Fig. 3D and fig. S9).

Li X., Mei Q., Yu Q., Wang M., He F., Xiao D., Liu H., Ge F., Yu X, & Li S. (2023). The TORC1 activates Rpd3L complex to deacetylate Ino80 and H2A.Z and repress autophagy. Science Advances, 9(10), eade8312.

Publication 2023

anti-IgG Antibodies Antibodies, Anti-Idiotypic Antibody Specificity Cell Extracts Goat Histone H2a Immunoblotting Immunoglobulins INO80 protein, human Lysine Mus Rabbits

Top products related to «Antibody Specificity»

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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.

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DAPI is a fluorescent dye that binds strongly to adenine-thymine (A-T) rich regions in DNA. It is commonly used as a nuclear counterstain in fluorescence microscopy to visualize and locate cell nuclei.

Triton x 100 by Merck Group

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Triton X-100 is a non-ionic surfactant commonly used in various laboratory applications. It functions as a detergent and solubilizing agent, facilitating the solubilization and extraction of proteins and other biomolecules from biological samples.

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Bovine serum albumin (BSA) is a common laboratory reagent derived from bovine blood plasma. It is a protein that serves as a stabilizer and blocking agent in various biochemical and immunological applications. BSA is widely used to maintain the activity and solubility of enzymes, proteins, and other biomolecules in experimental settings.

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Alexa Fluor 488 is a fluorescent dye used in various biotechnological applications. It has an excitation maximum at 495 nm and an emission maximum at 519 nm, producing a green fluorescent signal. Alexa Fluor 488 is known for its brightness, photostability, and pH-insensitivity, making it a popular choice for labeling biomolecules in biological research.

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The FACSCalibur is a flow cytometry system designed for multi-parameter analysis of cells and other particles. It features a blue (488 nm) and a red (635 nm) laser for excitation of fluorescent dyes. The instrument is capable of detecting forward scatter, side scatter, and up to four fluorescent parameters simultaneously.

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β-actin is a protein that is found in all eukaryotic cells and is involved in the structure and function of the cytoskeleton. It is a key component of the actin filaments that make up the cytoskeleton and plays a critical role in cell motility, cell division, and other cellular processes.

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What are the different factors that can influence antibody specificity?

Antibody specificity can be influenced by a variety of factors, including the structure and composition of the antigen, the immunization process, the antibody production methods, and the purification techniques used. Factors like the choice of adjuvant, the route of immunization, and the specifics of the antibody generation process can all impact the final specificity of the antibody. Understanding these variables is crucial for optimizing antibody specificity for your research needs.

How can PubCompare.ai help me enhance my antibody specificity research?

PubCompare.ai offers several key capabilities to streamline your antibody specificity research: 1. Screen protocol literature more efficiently by leveraging the platform's AI-powered search and analysis tools. 2. Identify critical insights that can help you pinpoint the most effective protocols for your specific research goals. 3. Utilize the platform's AI-driven comparisons to highlight key differences in protocol effectiveness, enabling you to choose the best option for reproducibility and accuracy. By empowering you to discover the most optimal antibody specificity protocols, PubCompare.ai can help you enhance your research outcomes and save time in the process.

What are some common challenges researchers face when working with antibody specificity?

Some of the key challenges researchers may encounter when working with antibody specificity include: 1. Cross-reactivity: Antibodies may bind to multiple antigens, leading to non-specific results. Optimizing specificity is crucial to avoid false positives. 2. Sensitivity: Achieving the right balance between specificity and sensitivity is often a delicate process. 3. Batch-to-batch variability: The specificity of antibodies can vary between production batches, requiring careful quality control. 4. Sample complexity: Complex biological samples can make it difficult to accurately measure and compare antibody specificity. PubCompare.ai can help address these challenges by providing insights and tools to streamline your antibody specificity research.

How does PubCompare.ai's AI-powered analysis help researchers identify the most effective antibody specificity protocols?

PubCompare.ai's AI-powered analysis helps researchers identify the most effective antibody specificity protocols in a few key ways: 1. Tha platform can rapidly screen a large volume of protocol literature, pre-prints, and patents to uncover the latest advancements and best practices. 2. Its AI-driven comparisons can highlight the critical differences between protocols, such as their impact on specificity, sensitivity, and reproducibility. 3. By surfacing these insights, PubCompare.ai empowers researchers to make more informed decisions about the optimal protocols for their specific research needs. This can lead to improved experimental design, more reliable results, and accelerated progress in your antibody specificity research.

More about "Antibody Specificity"

Antibody specificity is a crucial aspect of effective research, diagnosis, and treatment.
It refers to the degree to which an antibody recognizes and binds to a particular antigen.
Optimizing antibody specificity is essential for reliable and accurate results.
Antibody specificity is influenced by various factors, including the structure and composition of the antibody, the nature of the antigen, and the experimental conditions.
Techniques such as PVDF membranes, DAPI staining, and the use of detergents like Triton X-100 can help researchers assess and improve antibody specificity.
Bovine serum albumin (BSA) is commonly used as a blocking agent to reduce non-specific binding, while Alexa Fluor 488 is a popular fluorescent label used to visualize and quantify antibody binding.
Flow cytometry instruments like the FACSCalibur can also be employed to analyze antibody-antigen interactions.
To further enhance antibody specificity research, researchers can utilize tools like PubCompare.ai.
This AI-powered platform enables the discovery of the best protocols and products by analyzing literature, preprints, and patents.
By leveraging PubCompare.ai's insights, researchers can streamline their workflow, optimize their experimental design, and improve their results.
Additionally, researchers may encounter the need to detect and quantify proteins, such as β-actin, which is often used as a loading control in Western blotting.
The Pierce BCA Protein Assay Kit can be a valuable tool for accurately measuring protein concentrations in these experiments.
By incorporating these insights and techniques, researchers can enhance their understanding of antibody specificity and develop more reliable and effective research, diagnostic, and therapeutic strategies.