Customize GIS2 Antibody

Description

Introduction to GIS2

GIS2 (Gene Involved in Stress 2) is a yeast (Saccharomyces cerevisiae) protein implicated in mRNA handling during cellular stress. It is a zinc finger-containing protein that interacts with components of the translation machinery and localizes to cytoplasmic RNA-protein granules, including processing bodies (P-bodies) and stress granules . Human orthologs of GIS2 include CNBP (CCHC-type zinc finger nucleic acid-binding protein), which shares functional similarities in mRNA regulation .

Biological Functions of GIS2

Key roles of GIS2 in cellular processes:

mRNA Translation Regulation: GIS2 interacts with poly(A)-binding protein (Pab1) and translation initiation factors (eIF4G1/2) in an RNA-dependent manner, suggesting a role in modulating translation under stress .
Stress Granule Formation: GIS2 localizes to stress granules during arsenite-induced stress, indicating involvement in mRNA storage and repression during adverse conditions .
Proteomic Impact: Deletion of GIS2 in yeast reduces abundance of proteins like Imd2, Smd3, and Fap1, though its regulatory role under zinc deficiency appears limited .

Functional comparison of GIS2 and human CNBP:

Feature	Yeast GIS2	Human CNBP
RNA Binding	Binds mRNA via zinc finger motifs	Binds nucleic acids via CCHC domains
Stress Granule Localization	Yes	Yes (upon arsenite treatment)
Protein Interactions	Pab1, eIF4G1/2, ribosomal proteins	Poly(A)-binding protein

Antibody Validation and Use Cases

GIT2 Antibody (Proteintech 67858-1-Ig):
- Target: Human GIT2 (G protein-coupled receptor kinase-interacting ArfGAP 2) .
- Applications: Validated in Western blot (WB), immunohistochemistry (IHC), and ELISA .
- Reactivities: Human, rat .
- Dilution Recommendations:
  
  Application Dilution Range
  WB 1:5,000–1:50,000
  IHC 1:500–1:2,000

Application	Dilution Range
WB	1:5,000–1:50,000
IHC	1:500–1:2,000

Key Findings from GIS2-Related Studies

Yeast GIS2 Interactome:
- Interacts with translation initiation factors (eIF4G1/2) and ribosomal proteins, linking it to mRNA translation .
- Associates with stress granule components under stress conditions .
Human CNBP: Shares RNA-binding properties and stress granule localization with GIS2, suggesting conserved roles in mRNA regulation .

Challenges and Opportunities in GIS2 Research

Antibody Availability: No direct references to GIS2-specific antibodies were found in the provided materials. Research on GIS2 likely relies on epitope-tagged constructs or ortholog-targeting reagents (e.g., anti-CNBP antibodies).
Functional Redundancy: GIS2’s limited impact on the yeast proteome under zinc deficiency suggests overlapping roles with other regulators .

Future Directions

Development of GIS2-specific antibodies would enhance mechanistic studies of its role in stress responses.
Comparative studies between GIS2 and CNBP could elucidate conserved pathways in mRNA metabolism.

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

GIS2 antibody; At5g06650 antibody; Zinc finger protein GIS2 antibody; Protein GLABROUS INFLORESCENCE STEMS 2 antibody

Target Names

GIS2

Uniprot No.

Q9FG05

Target Background

Function

GIS2 Antibody targets a putative transcription factor believed to be essential for the initiation of trichomes on inflorescences in response to gibberellin and cytokinin. It does not play a role in regulating trichome branching. Functionally, GIS2 Antibody is equivalent to ZFP8 Antibody.

Database Links

KEGG: ath:AT5G06650

STRING: 3702.AT5G06650.1

Subcellular Location

Nucleus.

Tissue Specificity

Expressed in inflorescence meristems, floral meristems and stem epidermis.

Q&A

Here’s a structured FAQ collection for GIS2 antibody research, synthesized from peer-reviewed studies and technical guidelines:

Advanced Research Questions

How to resolve contradictions in GIS2 interaction data across studies?

Case analysis:

Observation	Study A	Study B	Resolution Strategy
eIF4G interaction	Strong in Co-IP	Weak in polyribosome fractions	Vary lysis buffers (RNase sensitivity)
Ribosomal association	Minimal (4.5%)	Significant (39.7% in monoribosomes)	Compare cycloheximide vs. untreated lysates

What methodologies confirm GIS2’s dual role in mRNA stability and translation?

Integrated workflow:
- mRNA decay assay: Use XRN1 knockout strains + RNA-seq to quantify stability of Gis2-bound transcripts.
- Ribosome profiling: Compare polysome loading in GIS2Δ vs. wild-type under stress .
- CRISPR-Cas9 knock-in: Tag endogenous GIS2 with GFP for live-cell imaging of granule dynamics.

How to design a study comparing yeast Gis2 and human CNBP orthologs?

Key considerations:
- Conserved domains: Align Zn-knuckle motifs (critical for RNA binding) using Clustal Omega.
- Functional complementation: Express human CNBP in GIS2Δ yeast and assay rescue of translation defects .
- Cross-species Co-IP: Use anti-CNBP antibodies (validated via siRNA knockdown) to test interactions with human eIF4G/PABPC1 .

Methodological Best Practices

Antibody Validation Table

Application	Validation Step	Acceptable Result
Western Blot	Lysate from GIS2Δ strain	No band at ~75 kDa
Immunoprecipitation	RNase-treated vs. untreated	Reduced eIF4G1 signal post-RNase
IHC (tissue)	siRNA knockdown + staining	≥70% signal reduction

Polyribosome Association Quantification

Condition	% Gis2 in Polyribosomes	Key Controls
+Cycloheximide	4.5%	Rpl1A/B blots
-Cycloheximide	1.3%	80S peak analysis
+Micrococcal nuclease	1.2%	mRNA degradation QC

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GIS2 Antibody