FABP7 Antibody, FITC conjugated

What is FABP7 and why is it significant in research applications?

FABP7 (also known as B-FABP or BLBP) is a member of the FABP family of lipid chaperones involved in the uptake and intracellular trafficking of fatty acids. This 15 kDa protein (observed in Western blots) has emerged as an important research target due to its differential expression in normal and pathological states . FABP7 is particularly significant in melanoma research, where it has been identified as a potential melanoma antigen involved in cell proliferation and invasion . For effective research applications, FITC-conjugated FABP7 antibodies allow direct visualization of expression patterns without requiring secondary antibody incubation steps, streamlining immunofluorescence protocols while maintaining sensitivity.

Which tissue types typically express FABP7?

FABP7 shows tissue-specific expression patterns that researchers should consider when designing experiments. According to immunohistochemical analyses, FABP7 protein has been positively detected in:

When using FITC-conjugated antibodies, researchers should consider autofluorescence backgrounds of these tissues and implement appropriate controls to distinguish specific from non-specific signals.

What are the optimal conditions for immunofluorescence detection using FITC-conjugated FABP7 antibodies?

While standard immunofluorescence protocols apply to FITC-conjugated antibodies, FABP7 detection requires specific optimization. Based on successful detection parameters for unconjugated FABP7 antibodies, researchers should:

• Perform antigen retrieval using TE buffer pH 9.0 for formalin-fixed paraffin-embedded tissues (alternatively, citrate buffer pH 6.0)

• Use antibody dilutions ranging from 1:50-1:500, optimizing for your specific tissue

• Include DAPI nuclear counterstain to facilitate cellular localization

• Implement photobleaching preventative measures (anti-fade mounting media, reduced exposure times)

• Image promptly after staining as FITC fluorescence degrades over time

For negative controls, use isotype-matched FITC-conjugated IgG to establish background fluorescence levels in your specific tissue.

How can researchers validate FABP7 antibody specificity for their experimental system?

Antibody validation is critical for reliable research outcomes. A comprehensive validation approach should include:

• Western blot analysis comparing expression across relevant tissues, confirming a single band at approximately 15 kDa (the observed molecular weight for FABP7)

• Parallel analysis using multiple antibody clones targeting different FABP7 epitopes

• Testing in known positive tissues (human brain tissue, melanoma) and negative controls

• Competitive binding assays using recombinant human FABP7 protein (Met1-Ala132)

• Testing in FABP7 knockout systems as generated by CRISPR/Cas9 in the C57BL/6 background

When publishing, researchers should report the antibody's RRID (Research Resource Identifier) - for example, AB_2100456 for certain commercial FABP7 antibodies .

How does FABP7 expression change during melanoma progression and what are the implications for experimental design?

FABP7 expression exhibits significant changes during melanoma progression that researchers must consider when designing longitudinal studies. Immunohistochemical assessment of 37 paired tissues of primary and metastatic melanomas revealed that FABP7 protein was detected in 27 of 37 (73%) primary melanomas but only in 10 of 37 (27%) metastatic melanomas (P<0.0001) . This differential expression pattern requires researchers to:

• Clearly identify and document tumor stage and classification

• Consider temporal aspects of FABP7 expression in experimental timelines

• Include multiple sampling timepoints in progression models

• Correlate FABP7 expression with clinical outcomes and prognostic indicators

The survival impact data provides compelling evidence for FABP7's biological significance:

These statistics emphasize FABP7's potential as a prognostic biomarker, warranting careful fluorescent quantification in clinical specimens .

What experimental approaches enable functional analysis of FABP7 in cancer models?

To determine FABP7's functional roles, researchers can manipulate its expression using several methodologies:

• siRNA-mediated knockdown: Specific small interfering RNAs have successfully down-regulated FABP7 expression, resulting in suppressed in vitro cell proliferation and Matrigel invasion in certain melanoma cell lines .

• Overexpression studies: Transfection with FABP7 cDNA in FABP7-negative cell lines (such as embryonic kidney 293T cells) has demonstrated enhanced cell proliferation and invasion, confirming FABP7's role in malignant phenotypes .

• CRISPR/Cas9 knockout models: Complete FABP7 gene deletion in mouse models has enabled assessment of phenotypic consequences, revealing roles in auditory function preservation and metabolic pathway modulation .

• Promoter activity analysis: Co-transfection with FABP7 promoter constructs (such as -1122+89 pGL4-FABP7) can elucidate transcriptional regulation mechanisms across different cell types .

When using FITC-conjugated antibodies in these models, researchers should quantify fluorescence intensity using standardized exposure settings and include appropriate controls for each experimental condition.

How can researchers address discrepancies between FABP7 protein detection by immunofluorescence versus other methods?

Methodological discrepancies are common challenges in FABP7 research. When immunofluorescence results using FITC-conjugated antibodies conflict with other detection methods, consider:

• Epitope accessibility issues: Different fixation protocols may affect antibody binding. Test multiple fixatives (4% PFA, methanol, acetone) and antigen retrieval methods.

• Expression threshold detection limits: Western blot detection of FABP7 has successfully identified the protein in human cerebellum and hippocampus tissues using specific conditions (0.2 mg/mL lysate concentration and 50 μg/mL antibody concentration) . Immunofluorescence may require higher antibody concentrations.

• Subcellular localization variations: FABP7 distribution may vary between cytoplasmic and nuclear compartments depending on cell state. Z-stack imaging may reveal localization patterns missed in single-plane microscopy.

• Post-translational modifications: These may affect epitope recognition differently across methods. Multiple antibodies targeting different regions can help resolve this issue.

What factors impact FABP7 detection in circulating tumor cells (CTCs) using fluorescent antibodies?

FABP7 has potential as a surrogate mRNA biomarker for circulating tumor cells in melanoma. When developing fluorescence-based detection protocols, researchers should consider:

• The sensitivity threshold required for rare cell detection (CTCs occur at approximately 1-10 cells/mL blood)

• Optimizing signal amplification techniques for low abundance targets

• Implementing multiparameter analysis to distinguish CTCs from normal blood cells

• Correlating FABP7 protein expression with validated CTC qRT assays for FABP7 mRNA detection

Researchers have successfully used FABP7 as a CTC biomarker in melanoma patients, suggesting FITC-conjugated antibodies could enable rapid screening with appropriate protocol optimization .

How should researchers interpret FABP7 expression in relation to metabolic pathways?

Studies of Fabp7 knockout mice reveal complex metabolic interactions that should inform research design. RNA sequencing showed that genes associated with the electron transport chain were upregulated or less impaired in Fabp7 KO mice . Additionally, metabolomic analysis revealed decreased glutamate and aspartate levels in these knockouts . When investigating FABP7's role:

• Consider parallel assessment of fatty acid metabolism markers

• Measure mitochondrial function parameters alongside FABP7 expression

• Evaluate glutamate/aspartate levels as potential downstream effectors

• Design experiments that assess both FABP7 expression and functional metabolic outcomes

FITC-conjugated antibodies can facilitate co-localization studies with metabolic markers when using appropriate filter sets to avoid spectral overlap.

What are the considerations for using FITC-conjugated FABP7 antibodies in multiplexed imaging approaches?

As imaging technologies advance, researchers increasingly implement multiplexed approaches to visualize multiple targets simultaneously. When incorporating FITC-conjugated FABP7 antibodies:

• Select complementary fluorophores with minimal spectral overlap (typically Cy3, Cy5, or APC)

• Account for FITC's relatively rapid photobleaching by imaging this channel first or using anti-fade reagents

• Establish rigorous controls for potential antibody cross-reactivity

• Consider sequential staining protocols if antibody host species conflicts arise

• Implement spectral unmixing algorithms for optimal signal separation

Successful FABP7 detection has been demonstrated in tissues with complex autofluorescence profiles like brain and adipose tissue, suggesting multiplexed approaches are feasible with proper controls .

How can FABP7 antibodies be used to investigate its role in neural development and pathology?

FABP7's expression in radial glial cells during development points to important neurobiological functions. Researchers investigating these roles should:

• Design developmental timecourse studies spanning critical neurodevelopmental windows

• Combine FITC-FABP7 staining with markers of neural progenitors, mature neurons, and glia

• Consider region-specific expression patterns within the CNS (cerebellum vs. hippocampus)

• Correlate protein expression with functional assays of neural progenitor proliferation and differentiation

Positive immunofluorescence has been detected in rat brain tissue, suggesting cross-species applicability of certain FABP7 antibodies for comparative developmental studies .