Customize BXL2 Antibody

Description

Definition and Biological Context

BCL2 antibodies are immunodetection tools designed to identify the BCL2 protein, a 26 kDa antiapoptotic molecule encoded by the BCL2 gene. BCL2 suppresses programmed cell death by inhibiting mitochondrial membrane permeabilization and cytochrome c release, thereby blocking caspase activation . These antibodies are pivotal in research and diagnostics for distinguishing normal and neoplastic tissues, particularly in lymphomas .

Targets and Isoforms

Primary target: BCL2α isoform (239 amino acids) .
Cross-reactivity: Most antibodies (e.g., clone 100, E17, SP66) do not bind Bcl-x or Bax .

Applications

Diagnostic: Differentiates reactive follicular hyperplasia from follicular lymphoma .
Research: Measures BCL2 expression in apoptosis studies, cancer prognosis, and drug resistance mechanisms .
Techniques: Immunohistochemistry (IHC), Western blot (WB), flow cytometry .

Technical Performance Variability

Antibody Clone	Epitope Specificity	Detection Rate in DLBCL	Key Features
124 (Mouse mAb)	C-terminal region	34% (32/94 cases)	Lower sensitivity in translocation/amplification-positive DLBCL
E17 (Rabbit mAb)	Mid-region	62% (58/94 cases)	Detects high antigen levels; useful in ABC-DLBCL
SP66 (Rabbit mAb)	N-terminal region	80% (75/94 cases)	Superior prognostic stratification in MYC+ DLBCL

Drug Resistance Mechanisms

Venetoclax Resistance: Hyperphosphorylation of BCL2 family proteins and epigenetic modifications enable evasion of BCL2 inhibitors like venetoclax .
Therapeutic Targeting: Clinical trials (e.g., NCT04575064) explore combinations with kinase inhibitors to overcome resistance .

Antibody Validation

NordiQC Assessments: Optimal IHC requires moderate-to-strong cytoplasmic staining in T-cells and mantle zones .
Cross-Platform Consistency: Clone SP66 outperforms 124 in detecting BCL2 amplification and translocations .

Emerging Directions

Biomarker Development: BCL2 mutations (e.g., G33R, A43T) enhance antiapoptotic activity and correlate with early FL transformation .
Multiplex Assays: Integration with MYC and TP53 profiling refines risk stratification in hematologic malignancies .

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

BXL2 antibody; At1g02640 antibody; T14P4.11 antibody; T14P4.8Probable beta-D-xylosidase 2 antibody; AtBXL2 antibody; EC 3.2.1.- antibody

Target Names

BXL2

Uniprot No.

Q94KD8

Target Background

Function

BXL2 Antibody may play a role in the remodeling of xylans during vascular development.

Database Links

KEGG: ath:AT1G02640

STRING: 3702.AT1G02640.1

UniGene: At.25322

Protein Families

Glycosyl hydrolase 3 family

Subcellular Location

Secreted, extracellular space, extracellular matrix.

Tissue Specificity

Expressed in leaves, stems, seedlings, roots, inflorescences and siliques.

Q&A

FAQs for BCL2 Antibody Research

Advanced Research Questions

How to resolve contradictory BCL2 expression data across studies?
- Epitope variability: Antibodies targeting different regions (e.g., N-terminal vs. C-terminal epitopes) may yield discordant results. For example, clone EPR17509 detects a 26 kDa band , while others might detect splice variants .
- Tissue-specific artifacts: Differences in fixation (e.g., formalin vs. frozen sections) impact epitope accessibility .
- Orthogonal validation: Combine IHC with RNA in situ hybridization (ISH) or CRISPR-Cas9 knockout models .
What experimental designs mitigate cross-reactivity in multiplex assays?
- Use antibodies with distinct labels (e.g., FITC vs. IRDye®800CW) .
- Pre-absorb secondary antibodies to minimize interspecies reactivity .
- Validate in knockout models to confirm target exclusivity .
How to analyze BCL2’s role in apoptosis using functional assays?
- Combine antibody-based detection (e.g., flow cytometry for BCL2 levels) with functional readouts like mitochondrial membrane potential assays .
- Use co-immunoprecipitation to study BCL2-Bax/Bak interactions .

Table 1: Comparison of BCL2 Antibody Clones

Clone	Epitope Specificity	Applications	Validation Method	Key Findings
EPR17509	Human BCL2 (native)	WB, IHC, Flow Cytometry	Knockout cell lysates, isotype controls	Detects 26 kDa band; no cross-reactivity
10C4	Mouse/Rat BCL2	Intracellular Flow Cytometry	Splenocyte/thymocyte staining	Cross-reacts with rat BCL2
SP66	Human BCL2	IHC, ISH	DLBCL tissue microarrays	Higher sensitivity in follicular lymphoma

Methodological Considerations

Multiplex IHC: Use enzyme-labeled antibodies (e.g., HRP/AP) with sequential staining to avoid signal overlap .
Quantitative Analysis: Normalize BCL2 levels to housekeeping proteins (e.g., GAPDH) in Western blots .
Data Reproducibility: Document antigen retrieval conditions (e.g., pH 9.0 buffer for 20 minutes) and antibody lot numbers .

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BXL2 Antibody