EEF1G Antibody

Introduction to EEF1G Antibody

EEF1G antibodies are immunological reagents specifically designed to recognize and bind to the eukaryotic translation elongation factor 1 gamma protein. These antibodies are developed for the immunodetection of EEF1G, which functions as a subunit of the elongation factor-1 complex responsible for the enzymatic delivery of aminoacyl tRNAs to the ribosome during protein synthesis . In humans, the canonical EEF1G protein consists of 437 amino acid residues with an approximate molecular weight of 50.1 kDa .

The target protein recognized by these antibodies, EEF1G, contains an N-terminal glutathione transferase domain that likely plays a role in regulating the assembly of multisubunit complexes containing this elongation factor and aminoacyl-tRNA synthetases . This protein is also known by several synonyms, including GIG35, EF-1-gamma, PRO1608, eEF-1B gamma, and EF1G .

Types and Origins

EEF1G antibodies are available in multiple formats to accommodate various research needs. They are primarily categorized as either monoclonal or polyclonal antibodies:

1. Monoclonal Antibodies: These are highly specific antibodies derived from a single B-cell clone. For example, the Proteintech mouse monoclonal EEF1G antibody (clone 1G7C6) is generated with recombinant protein of human EEF1G .

2. Polyclonal Antibodies: These antibodies are derived from multiple B-cell lineages. The Assay Genie EEF1G rabbit polyclonal antibody (CAB7891) is produced using a recombinant fusion protein containing a sequence corresponding to amino acids 168-437 of human EEF1G .

Applications of EEF1G Antibodies in Research

EEF1G antibodies serve as versatile tools across various experimental techniques in molecular and cellular biology research.

Validated Applications

The primary applications of EEF1G antibodies include:

1. Western Blot (WB): EEF1G antibodies effectively detect the target protein at approximately 50 kDa in denatured protein samples .

2. Immunocytochemistry/Immunofluorescence (IF/ICC): These antibodies can visualize the intracellular localization of EEF1G, which is predominantly cytoplasmic in distribution .

3. Enzyme-Linked Immunosorbent Assay (ELISA): EEF1G antibodies can be utilized in ELISA-based detection methods for quantitative analysis .

4. Immunohistochemistry (IHC): Particularly with polyclonal antibodies, EEF1G can be detected in tissue sections, enabling the study of its expression patterns across different tissues .

Target Expression Patterns

EEF1G antibodies have revealed differential expression patterns across various tissues:

• High expression in pancreatic tumor tissue

• Moderate expression in normal kidney, intestine, pancreas, stomach, lung, brain, spleen, and liver tissues

• Upregulated expression in lung adenocarcinoma tissues compared to adjacent normal tissues

Normal Physiological Role

EEF1G functions as a component of the eukaryotic elongation factor-1 complex, which plays a central role in protein synthesis. Specifically:

1. The eEF1 complex consists of two functional parts: eEF1A and the eEF1B complex .

2. EEF1G is a subunit of the eEF1B complex, which also includes eEF1B2 and eEF1D .

3. This complex possesses guanine nucleotide exchange activity that activates eEF1A, which delivers aminoacyl-tRNAs to the ribosome during translation .

4. The C-terminal region of EEF1G appears functionally important for maintaining eEF1B2 and eEF1D expression .

Role in Viral Replication

Research using EEF1G antibodies has revealed significant roles of this protein in viral replication:

1. Influenza A Virus: Studies indicate that EEF1G contributes to influenza A virus replication in a strain-specific manner .

2. HIV-1 Replication: EEF1G has been identified as a critical HIV-1 reverse transcription cofactor:

   • It coimmunoprecipitates with HIV-1 reverse transcriptase (RT) and integrase (IN) .

   • EEF1G associates with reverse transcription complexes (RTCs) in infected cells .

   • Complete loss of reverse transcription stimulatory activity was observed when EEF1G levels were reduced by 70-90% .

The following table summarizes experimental findings on EEF1G's role in viral replication:

Oncological Significance

Recent research using EEF1G antibodies has revealed important implications in cancer biology:

Recommended Dilutions

Proper antibody dilution is crucial for optimal results across different applications:

Table 3: Recommended dilutions for EEF1G antibodies by application

Detection Specificity

The specificity of EEF1G antibodies can be influenced by several factors:

1. Antibody Type: Monoclonal antibodies typically offer higher specificity but may recognize a single epitope, whereas polyclonal antibodies recognize multiple epitopes but might show more cross-reactivity .

2. Target Region: Some antibodies target specific regions of the EEF1G protein. For example, the rabbit monoclonal antibody used in certain studies detects a peptide distant from amino acids 182-186, making it suitable for detecting specific mutations in these regions .

3. Cross-Reactivity: While many EEF1G antibodies are validated for human samples, some also show reactivity with other species such as pig, rabbit, mouse, and rat .