Customize spr-3 Antibody

Description

Molecular and Functional Characteristics of SPTBN2/2887R Antibody

Target: Human Spectrin beta 3 (SPTBN2), a cytoskeletal protein critical for membrane stability and intracellular signaling.

Property	Details
Immunogen	Recombinant human Spectrin beta 3 fragment (aa356-475; proprietary sequence)
Host Species	Rabbit
Clonality	Monoclonal (IgG isotype)
Concentration	0.2 mg/ml (with BSA/azide); 1.0 mg/ml (azide-free formulation)
Applications	Western Blot, Flow Cytometry, Immunofluorescence, Immunohistochemistry (paraffin-embedded)
Theoretical MW	246 kDa (observed MW may vary due to post-translational modifications)

Western Blot Analysis

Detects SPTBN2 in PANC-1 cell lysates at ~246 kDa (predicted size) .
Key validation: Specific binding confirmed using recombinant human protein controls.

Immunofluorescence

Localizes SPTBN2 to the cell membrane and cytosolic compartments in HeLa cells .
Protocol: Fixed cells stained with SPTBN2/2887R (1:50 dilution) and CF488-conjugated secondary antibody.

Flow Cytometry

Surface expression analysis in HeLa cells showed distinct fluorescence shifts compared to isotype controls .
Gating strategy: Live-cell populations analyzed with forward/side scatter to exclude debris.

Comparative Advantages Over Polyclonal Antibodies

Feature	SPTBN2/2887R (Monoclonal)	Polyclonal Antibodies
Specificity	High (single epitope targeting)	Moderate (multiple epitopes)
Batch Consistency	Excellent	Variable
Background Noise	Low	Higher due to cross-reactivity

Technical Notes for Experimental Use

Storage:
- With azide: 2–8°C
- Azide-free: -20°C to -80°C
Recommended dilutions:
- Western Blot: 1:500–1:2,000
- Immunofluorescence: 1:50–1:200

Role in Immunotherapy and Disease Research

While not directly linked to immunotherapy, Spectrin beta 3 is implicated in cancer progression and neuronal disorders. Surface Plasmon Resonance (SPR)-based assays (as described for other antibodies ) could enhance functional studies of SPTBN2/2887R by:

Quantifying binding kinetics (e.g., $K_D$ ) to Spectrin beta 3 isoforms.
Screening for cross-reactivity with homologous proteins (e.g., Spectrin beta 1/2) .

Limitations and Future Directions

Epitope mapping: The exact binding region (aa356-475) is proprietary, limiting mechanistic studies .
Therapeutic potential: No clinical trials involving this antibody have been reported as of March 2025.

Product Specs

Buffer

Preservative: 0.03% Proclin 300
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

spr-3 antibody; C07A12.5 antibody; Suppressor of presenilin protein 3 antibody

Target Names

spr-3

Uniprot No.

Q17768

Target Background

Function

SPR-3 is a probable transcriptional regulator that participates in the transcriptional repression of the presenilin protein HOP-1.

Database Links

KEGG: cel:CELE_C07A12.5

STRING: 6239.C07A12.5a

UniGene: Cel.24627

Subcellular Location

Nucleus.

Q&A

Here’s a structured FAQ document for spr-3 Antibody research, prioritizing academic rigor, experimental methodology, and critical analysis. Content is synthesized from peer-reviewed protocols, patents, and SPR/antibody engineering literature:

Advanced Research Questions

How to resolve discrepancies between SPR and ELISA binding data for spr-3?

Factor	SPR	ELISA
Binding context	Solution-phase kinetics	Solid-phase, avidity effects
Data interpretation	Direct K<sub>D</sub> calculation	Semi-quantitative (relative OD)

Action: Perform steady-state SPR at higher analyte concentrations to detect weak interactions missed by ELISA .

What engineering strategies improve spr-3’s pH-dependent binding?

Framework humanization:
- Use VH3-07/JH4 frameworks to maintain CDR conformation while enhancing solubility .
- Validate mutations via alanine scanning of CDR-H3 (SEQ ID NO: 3) .
Consensus sequence integration:
- Integrate motifs like CISPRGC to enhance FcRn engagement without altering specificity .

How to design a spr-3 variant with reduced non-specific binding?

Approach:
- Screen spr-3 against a non-target protein panel (e.g., serum albumin, IgG) via SPR .
- Introduce electrostatic steering mutations (e.g., D→K substitutions in CDR-L1) to destabilize off-target interactions .
- Validate using cross-linking MS to identify residual binding partners .

Data Contradiction Analysis

Conflicting neutralization results between FRNT and SPR: How to interpret?

Root causes:
- Epitope accessibility: SPR measures 1:1 binding, while FRNT requires bivalent engagement .
- Solution vs. cell surface: SPR uses purified receptor; FRNT includes membrane topology effects .
Resolution: Perform Fab fragment SPR to simulate monovalent binding and compare with intact IgG FRNT .

Key Experimental Parameters from Literature

Assay	Critical Parameter	Optimal Value
SPR kinetics	Immobilization level	50–100 RU
Competition ELISA	EDIII concentration range	0.1–1000 nM
Humanization	VH framework identity	≥85% (VH3-07/JH4)

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spr-3 Antibody