Recombinant Human PiggyBac transposable element-derived protein 2 (PGBD2)

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Cat. No.

BT1177899

Source

Yeast

Synonyms

PGBD2; PiggyBac transposable element-derived protein 2

Purity

>85% (SDS-PAGE)

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Cat. No.

BT1177899

Source

E.coli

Synonyms

PGBD2; PiggyBac transposable element-derived protein 2

Purity

>85% (SDS-PAGE)

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Cat. No.

BT1177899

Source

E.coli

Synonyms

PGBD2; PiggyBac transposable element-derived protein 2

Purity

>85% (SDS-PAGE)

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Cat. No.

BT1177899

Source

Baculovirus

Synonyms

PGBD2; PiggyBac transposable element-derived protein 2

Purity

>85% (SDS-PAGE)

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Cat. No.

BT1177899

Source

Mammalian cell

Synonyms

PGBD2; PiggyBac transposable element-derived protein 2

Purity

>85% (SDS-PAGE)

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Description

Gene and Protein Overview

The PGBD2 gene (NCBI Gene ID: 267002) encodes a protein with homology to transposases from the piggyBac superfamily . Key features include:

Property	Detail
Chromosomal Location	Chromosome 1 (GRCh38.p14)
Protein Accession	NP_001017434.1 (isoform 1), Q6P3X8 (UniProt)
Domains	Conserved DDE/D catalytic triad (aspartic acid residues)
Expression	Detected in brain, thymus, and thyroid tissues

Research Applications and Challenges

While recombinant PGBD2 has not been extensively characterized, insights from homologous proteins suggest potential uses:

Table 1: Potential Applications

Application	Rationale
Genome Editing	TTAA-specific integration for precise gene insertion
Developmental Biology	Roles in neurogenesis inferred from expression in brain tissues
Cancer Research	CRISPR screens link PGBD2 to cell viability in glioblastoma models

Challenges:

Low Annotation: PGBD2 is classified as understudied, with limited functional data (Pharos score: 0.35) .
Technical Barriers: No published protocols for recombinant PGBD2 purification or activity assays.

Comparative Analysis with PGBD5

Studies on the closely related PGBD5 provide a framework for understanding PGBD2:

Feature	PGBD2	PGBD5
Transposase Activity	Predicted but unconfirmed	Confirmed in human cells
Expression	Broad tissue distribution	Enriched in neurons and embryos
Conservation	Vertebrate-specific	Deeply conserved across vertebrates

Future Directions

Key research priorities include:

Functional Validation: Developing assays to test recombinant PGBD2’s DNA transposition activity.
Structural Studies: Resolving 3D structures to clarify catalytic residues and substrate binding.
Therapeutic Exploration: Investigating roles in somatic genome diversification or disease.

Product Specs

Form

Lyophilized powder
Note: While we prioritize shipping the format currently in stock, please specify your format preference during order placement for customized preparation.

Lead Time

Delivery times vary depending on the purchasing method and location. Please contact your local distributor for precise delivery estimates.
Note: Standard shipping includes blue ice packs. Dry ice shipping requires advance notice and incurs additional charges.

Notes

Avoid repeated freeze-thaw cycles. Store working aliquots at 4°C for up to one week.

Reconstitution

Centrifuge the vial briefly before opening to collect the contents. Reconstitute the protein in sterile deionized water to a concentration of 0.1-1.0 mg/mL. For long-term storage, we recommend adding 5-50% glycerol (final concentration) and aliquoting at -20°C/-80°C. Our standard glycerol concentration is 50%, which can serve as a reference.

Shelf Life

Shelf life depends on various factors, including storage conditions, buffer composition, temperature, and the protein's inherent stability. Generally, liquid formulations have a 6-month shelf life at -20°C/-80°C, while lyophilized forms have a 12-month shelf life at -20°C/-80°C.

Storage Condition

Upon receipt, store at -20°C/-80°C. Aliquot for multiple uses to prevent repeated freeze-thaw cycles.

Tag Info

Tag type is determined during the manufacturing process.
The tag type will be determined during production. If you require a specific tag, please inform us, and we will prioritize its development.

Synonyms

PGBD2; PiggyBac transposable element-derived protein 2

Buffer Before Lyophilization

Tris/PBS-based buffer, 6% Trehalose.

Datasheet

Please contact us to get it.

Expression Region

1-592

Protein Length

full length protein

Purity

>85% (SDS-PAGE)

Species

Homo sapiens (Human)

Target Names

PGBD2

Target Protein Sequence

MASTSRDVIA GRGIHSKVKS AKLLEVLNAM EEEESNNNRE EIFIAPPDNA AGEFTDEDSG DEDSQRGAHL PGSVLHASVL CEDSGTGEDN DDLELQPAKK RQKAVVKPQR IWTKRDIRPD FGSWTASDPH IEDLKSQELS PVGLFELFFD EGTINFIVNE TNRYAWQKNV NLSLTAQELK CVLGILILSG YISYPRRRMF WETSPDSHHH LVADAIRRDR FELIFSYLHF ADNNELDASD RFAKVRPLII RMNCNFQKHA PLEEFYSFGE SMCEYFGHRG SKQLHRGKPV RLGYKIWCGT TSRGYLVWFE PSQGTLFTKP DRSLDLGGSM VIKFVDALQE RGFLPYHIFF DKVFTSVKLM SILRKKGVKA TGTVREYRTE RCPLKDPKEL KKMKRGSFDY KVDESEEIIV CRWHDSSVVN ICSNAVGIEP VRLTSRHSGA AKTRTQVHQP SLVKLYQEKV GGVGRMDQNI AKYKVKIRGM KWYSSFIGYV IDAALNNAWQ LHRICCQDAQ VDLLAFRRYI ACVYLESNAD TTSQGRRSRR LETESRFDMI GHWIIHQDKR TRCALCHSQT NTRCEKCQKG VHAKCFREYH IR

Uniprot No.

Q6P3X8

Q&A

Basic Research Questions

How to validate recombinant PGBD2 protein functionality in mammalian cell assays?

Method: Use excision assays with plasmid-based piggyBac transposons. Co-transfect cells with recombinant PGBD2 and a reporter plasmid (e.g., pTpB with GFP/antibiotic resistance). Perform PCR amplification of excised transposons and Sanger sequencing to confirm TTAA target site reconstitution .
Controls: Include piggyBac transposase (positive control) and a catalytically inactive mutant (negative control). Quantify colony formation under antibiotic selection to assess transposition efficiency .

What are optimal Western blot conditions for detecting recombinant PGBD2?

Protocol:
- Primary antibody: Rabbit polyclonal anti-PGBD2 (1:500–1:2000 dilution) .
- Sample preparation: Use RIPA buffer with protease inhibitors. Load 20–30 µg of lysate per lane.
- Validation: Include recombinant PGBD2 control fragment (aa 71-144) for specificity verification .

Parameter	Recommendation
Expected MW	68 kDa (isoform 1)
Observed MW	May vary due to PTMs
Blocking Buffer	5% non-fat milk in TBST

How to resolve discrepancies in PGBD2 activity across experimental models?

Troubleshooting steps:
- Confirm protein expression via Western blot and GFP fluorescence .
- Optimize transfection ratios (e.g., 1:3 transposon:transposase plasmid) .
- Test truncated inverted terminal repeats (ITRs) to enhance sensitivity .

Advanced Research Questions

What structural features influence PGBD2’s transposase activity compared to other piggyBac proteins?

Key domains:
- Catalytic triad: DDD motif critical for DNA cleavage .
- Terminal inverted repeats (TIRs): 5′-CCGG-3′ conserved motif; ITR length (4–24 bp) affects binding specificity .
Functional assay: Perform EMSA with fluorescently labeled ITRs to compare binding affinity between PGBD2 and piggyBac transposase .

How to interpret conflicting data on PGBD2’s role in genomic instability?

Case study: Henssen et al. (2016) reported PGBD5-mediated transposition in cancer cells, but follow-up studies found no activity for PGBD2/PGBD5 .
Resolution strategy:
- Use qPCR to quantify transposon copy numbers post-transfection .
- Apply single-cell sequencing to detect rare integration events.

Factor	Impact on Activity
ITR truncation	↑ Sensitivity
Transposase dosage	↓ Overproduction inhibition
Host cell type	Varies (e.g., HEK293 vs. HT-1080)

What orthogonal methods confirm PGBD2’s interaction with chromatin?

Approach:
- CUT&Tag sequencing: Profile genome-wide binding sites using anti-PGBD2 antibodies .
- Co-immunoprecipitation: Identify protein partners (e.g., histone modifiers) in nuclear lysates .
- Live-cell imaging: Fuse PGBD2 with mNeonGreen to track subcellular localization .

Methodological Considerations

Protein storage: Aliquot recombinant PGBD2 in 50% glycerol buffer; avoid >3 freeze-thaw cycles .
Ethical guidelines: Adhere to NIH guidelines for recombinant DNA (Appendix M) when engineering transposon systems .

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Recombinant Human PiggyBac transposable element-derived protein 2 (PGBD2)

Description

Gene and Protein Overview

Research Applications and Challenges

Table 1: Potential Applications

Challenges:

Comparative Analysis with PGBD5

Future Directions

Product Specs

Q&A

Basic Research Questions

How to validate recombinant PGBD2 protein functionality in mammalian cell assays?

What are optimal Western blot conditions for detecting recombinant PGBD2?

How to resolve discrepancies in PGBD2 activity across experimental models?

Advanced Research Questions

What structural features influence PGBD2’s transposase activity compared to other piggyBac proteins?

How to interpret conflicting data on PGBD2’s role in genomic instability?

What orthogonal methods confirm PGBD2’s interaction with chromatin?

Methodological Considerations

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