OFUT36 Antibody

What is OFUT36 Antibody and what are its primary applications in plant research?

OFUT36 Antibody (Product Code: CSB-PA952094XA01DOA) is a polyclonal antibody raised in rabbits against recombinant Arabidopsis thaliana OFUT36 protein. This antibody is primarily utilized for ELISA and Western Blot applications to detect and quantify OFUT36 expression in plant tissues .

Methodological approach: For optimal results in Western blotting, use protein samples prepared in reducing conditions with proper loading controls. The antibody is suitable for detecting target proteins within experimental systems and should be validated for specific applications before use in critical studies.

What are the recommended storage conditions for OFUT36 Antibody to maintain its functionality?

The OFUT36 Antibody should be stored at -20°C or -80°C upon receipt, and repeated freeze-thaw cycles should be avoided to maintain antibody integrity and binding capacity .

Methodological approach: Aliquot the antibody into smaller volumes upon first thaw to minimize freeze-thaw cycles. When removing from storage, thaw the antibody on ice and centrifuge briefly before use to collect all material at the bottom of the tube. Return unused portions to -20°C or -80°C immediately after use.

What are the key specifications of the OFUT36 Antibody that researchers should be aware of?

The OFUT36 Antibody has the following specifications:

• Product Code: CSB-PA952094XA01DOA

• UniProt No.: Q9FK30

• Immunogen: Recombinant Arabidopsis thaliana OFUT36 protein

• Host Species: Rabbit

• Species Reactivity: Arabidopsis thaliana

• Tested Applications: ELISA, WB

• Format: Liquid

• Conjugate: Non-conjugated

• Storage Buffer: 0.03% Proclin 300, 50% Glycerol, 0.01M PBS, pH 7.4

• Purification Method: Antigen Affinity Purified

• Isotype: IgG

• Clonality: Polyclonal

How should researchers validate the specificity of OFUT36 Antibody?

Methodological approach: Researchers should implement multiple validation strategies to confirm antibody specificity:

• Positive and negative controls: Include known positive samples (expressing OFUT36) and negative controls (tissues or cells without OFUT36 expression)

• Western blot analysis: Verify single band of expected molecular weight

• Blocking peptide competition: Pre-incubate antibody with blocking peptide to confirm specific binding

• Secondary antibody controls: Run parallel samples with secondary antibody only

• Genetic validation: When possible, use OFUT36 knockout/knockdown samples

It's critical to remember that antibodies may produce false-positive signals, as highlighted in study regarding other antibodies, necessitating rigorous validation protocols.

What experimental conditions should be optimized when using OFUT36 Antibody for Western blot analysis of plant proteins?

Methodological approach: To optimize Western blot conditions specifically for plant proteins with OFUT36 Antibody:

• Sample preparation:

  • Include plant-specific protease inhibitors in extraction buffer

  • Consider using specialized plant protein extraction buffers to contend with polyphenols, polysaccharides, and other plant components

  • Optimize protein loading (typically 20-50 μg total protein)

• Blocking and antibody incubation:

  • Test different blocking solutions (5% BSA or milk in TBST)

  • Optimize primary antibody concentration (start with 1:1000 dilution)

  • Incubate primary antibody overnight at 4°C

  • Extend washing steps to minimize background common in plant samples

• Detection strategy:

  • Consider HRP-conjugated secondary antibodies with chemiluminescent detection

  • For quantitative analysis, use fluorescent secondary antibodies

• Controls:

  • Include recombinant OFUT36 protein as positive control

  • Include samples from OFUT36 knockout plants if available

How can researchers distinguish between specific and non-specific signals when using OFUT36 Antibody?

This is particularly important as false-positive antibody signals can lead to erroneous data and misinterpretations, as demonstrated in studies with other antibodies .

Methodological approach:

• Conduct parallel Western blot analyses with both reducing and non-reducing conditions to verify target band identification

• Perform pre-absorption tests by incubating the antibody with excess antigen before use

• Analyze samples from different tissues/developmental stages to establish expression patterns

• Implement peptide competition assays to confirm specificity

• Compare results with mRNA expression data using RT-PCR or RNA-seq

• Consider using a second antibody recognizing a different epitope on the same protein

• Always include appropriate positive and negative controls

Researchers should be aware that some antibodies that show non-specific signals in immunofluorescence also often show additional non-specific bands in Western blots .

What strategies can be employed to improve OFUT36 Antibody performance in challenging plant tissue samples?

Methodological approach:

• Tissue-specific optimization:

  • Modify protein extraction protocols based on tissue type (roots, leaves, flowers, etc.)

  • Consider specialized extraction buffers for tissues with high phenolic compounds

• Signal enhancement techniques:

  • Use signal amplification systems (e.g., biotin-streptavidin)

  • Implement tyramide signal amplification for low-abundance proteins

  • Try extended antibody incubation times (up to 48 hours at 4°C)

• Background reduction strategies:

  • Pre-absorb antibody with non-specific plant proteins

  • Use highly purified BSA for blocking

  • Include additional washing steps with higher detergent concentrations

  • Consider using plant-specific blocking reagents

• Cross-reactivity minimization:

  • Increase antibody dilution to reduce non-specific binding

  • Use more stringent washing conditions

  • Implement additional purification steps for the antibody

How should researchers design experiments to investigate OFUT36 protein function using this antibody?

Methodological approach:

• Experimental design framework:

  • Begin with expression profiling across tissues and developmental stages

  • Correlate protein levels with gene expression data

  • Design experiments to test protein response to relevant stimuli or stresses

  • Include appropriate physiological and molecular readouts

• Functional studies approach:

  • Couple antibody-based detection with genetic approaches (knockouts, overexpression)

  • Use co-immunoprecipitation with OFUT36 Antibody to identify interaction partners

  • Employ subcellular fractionation followed by Western blotting to determine localization

  • Consider chromatin immunoprecipitation if nuclear function is suspected

• Validation strategy:

  • Compare results from antibody-based techniques with other methodologies

  • Implement complementary approaches (e.g., tagged versions of OFUT36)

  • Confirm findings across multiple experimental systems

What are the considerations for using OFUT36 Antibody in immunoprecipitation experiments?

Methodological approach:

• Pre-clearing protocol:

  • Pre-clear lysates with Protein A/G beads to remove proteins that bind non-specifically

  • Include control IgG from the same species (rabbit) in parallel experiments

• Optimization parameters:

  • Test different antibody amounts (typically 1-5 μg per 1 mg of total protein)

  • Optimize incubation conditions (time, temperature, buffer composition)

  • Determine optimal bead type and amount (Protein A, Protein G, or combination)

• Washing stringency:

  • Implement a gradient of washing stringency to determine optimal conditions

  • Consider including detergents appropriate for plant membrane proteins if relevant

• Elution conditions:

  • Test different elution methods (low pH, high pH, competitive elution)

  • Optimize elution buffer composition to maintain protein activity if downstream functional assays are planned

How can researchers accurately quantify OFUT36 protein levels using this antibody in ELISA?

Methodological approach:

• ELISA protocol optimization:

  • Determine optimal antibody concentration through titration experiments

  • Test different coating buffers for maximal antigen binding

  • Optimize blocking conditions to minimize background

  • Determine ideal sample dilutions through preliminary experiments

• Quantification strategy:

  • Develop standard curves using recombinant OFUT36 protein

  • Implement appropriate statistical methods for data analysis

  • Use technical and biological replicates to ensure reproducibility

  • Consider including spike recovery tests to validate extraction efficiency

• Data interpretation:

  • Normalize results to total protein concentration or appropriate reference proteins

  • Account for matrix effects when analyzing different tissue types

  • Implement appropriate controls for each experiment

What techniques can researchers employ to troubleshoot inconsistent results with OFUT36 Antibody?

Methodological approach:

• Systematic analysis of variables:

  • Verify antibody quality through SDS-PAGE analysis

  • Test different lots of antibody if available

  • Examine all buffers and reagents for contamination or degradation

  • Analyze sample preparation protocols for potential issues

• Technical modifications:

  • Adjust antibody concentration (both higher and lower)

  • Modify incubation conditions (time, temperature)

  • Test different detection systems

  • Implement alternative blocking reagents

• Sample-related troubleshooting:

  • Examine protein extraction efficiency

  • Check for presence of interfering compounds

  • Verify protein integrity through total protein stains

  • Consider enrichment approaches for low-abundance targets

• Documentation and analysis:

  • Maintain detailed records of all experimental parameters

  • Implement statistical approaches to identify significant variables

  • Compare results with published literature or alternative methods