Customize TIM21 Antibody

Description

Antibody Characteristics

TIM21 antibodies are polyclonal rabbit IgG reagents validated for diverse applications. Key properties include:

Property	Details
Target Protein	Mitochondrial Import Inner Membrane Translocase Subunit Tim21 (UniProt: Q9BVV7)
Immunogen	Synthetic peptides or fusion proteins derived from human C18orf55
Reactivity	Human, Mouse, Rat
Applications	Western Blot (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), ELISA
Dilution Range	WB: 1:500–2,000; IHC: 1:50–1:500
Storage	-20°C to -80°C in PBS/glycerol buffers; avoid freeze-thaw cycles

Protein Localization Studies

Detects endogenous TIM21 in mitochondrial membranes via WB and IHC .
Used to study TIM21’s interaction with Tim50 and Tim23 in mitochondrial protein import .

Mechanistic Insights

Signal Recognition: TIM21 dissociates from Tim50 upon presequence peptide binding, enabling translocase reorganization .
Structural Dependencies: Tim23’s receptor domain is required for TIM21-Tim50 interaction in organello .

Key Research Findings

Functional Role: TIM21 is essential for recruiting respiratory chain assembly factors to the TIM23 complex .
Crosslinking Studies: Anti-TIM21 antibodies identified adducts with Tim50, confirming their proximity in mitochondrial membranes .
Disease Relevance: Dysregulation linked to mitochondrial disorders, though direct mechanisms remain under investigation .

Validation and Quality Control

Specificity: Verified via protein arrays (e.g., Novus Biologicals’ antibody tested against 384 non-target proteins) .
Citations: Proteintech’s 16641-1-AP is cited in 2 WB and 1 IF publication .

Product Specs

Buffer

Preservative: 0.03% ProClin 300; Constituents: 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

TIM21 antibody; SD3 antibody; At4g00026 antibody; F6N15 antibody; Probable mitochondrial import inner membrane translocase subunit TIM21 antibody; Protein SEGREGATION DISTORTION 3 antibody

Target Names

TIM21

Uniprot No.

Q1G3L1

Target Background

Function

This antibody targets a protein that may participate in the translocation of proteins containing transit peptides across the mitochondrial inner membrane.

Gene References Into Functions

Studies suggest a link between this protein and mitochondrial function. For example:

Increased expression of respiratory-chain subunit genes, leading to elevated intracellular ATP levels, has been observed following SD3 induction. (PMID: 22131050)

Database Links

KEGG: ath:AT4G00026

STRING: 3702.AT4G00026.1

UniGene: At.50764

Protein Families

TIM21 family

Subcellular Location

Mitochondrion inner membrane; Single-pass membrane protein.

Tissue Specificity

Expressed in leaves, sheath, flowers and buds. Lower expression in roots and stems.

Q&A

Here’s a structured collection of FAQs tailored to academic research scenarios involving TIM21 antibodies, synthesized from scientific literature and technical specifications:

Advanced Research Questions

How do I resolve contradictions in TIM21’s role in respiratory chain complex assembly?

Hypothesis Testing:
- Mitochondrial Fractionation: Compare TIM21 distribution in mitoplasts (inner membrane) vs. matrix fractions under varying OXPHOS conditions .
- Functional Rescue: Transfect TIMM21-deficient cells with wild-type vs. mutants (e.g., ΔN-terminal) and assess Complex IV activity via BN-PAGE .

Parameter	Method	Expected Outcome
Complex IV Assembly	Blue Native PAGE	Reduced assembly in TIMM21-KO cells
Protein Import	Radiolabeled precursor uptake	Delayed import in ΔN-TIMM21 mutants

What strategies enhance TIM21 antibody performance in low-abundance mitochondrial proteomics?

Optimization:
- Signal Amplification: Tyramide-based amplification for IHC .
- Crosslinking MS: Combine immunoprecipitation with crosslinking mass spectrometry (e.g., BioID proximity labeling) to map TIM21 interactomes .

How do I address conflicting reports on TIM21’s involvement in mitophagy?

Experimental Design:
- Stress Conditions: Induce mitophagy (e.g., CCCP treatment) and monitor TIM21-Parkin colocalization via confocal microscopy .
- Knockdown Validation: Use TIMM21 siRNA alongside ATG5/ATG7 knockouts to isolate mitophagy-specific effects .

Methodological Considerations

What statistical approaches are appropriate for TIM21-dependent mitochondrial import assays?

Analysis:
- Time-Course Data: Fit nonlinear regression curves to precursor import kinetics (GraphPad Prism).
- Multivariate Testing: Apply ANOVA with post-hoc Tukey tests when comparing multiple TIMM21 mutants .

How do I integrate TIM21 antibody data with multi-omics datasets (e.g., transcriptomics)?

Workflow:
- Correlation Analysis: Pair IHC scores (H-score) with RNA-seq data from the same tissue .
- Pathway Enrichment: Use tools like GSEA to link TIMM21 expression to OXPHOS or mitochondrial biogenesis pathways .

Technical Troubleshooting

Issue	Root Cause	Solution
Weak WB signal	Over-fixation in IHC	Reduce paraformaldehyde concentration to 2%
Non-specific IHC staining	Endogenous peroxidases	Quench with 3% H₂O₂ in methanol for 15 min

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TIM21 Antibody