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GADD45A cell lysate

Human GADD45A Baculovirus-Insect cells Overexpression Lysate

Gene Names
GADD45A; DDIT1; GADD45
Applications
Western Blot
Synonyms
GADD45A; Human GADD45A Baculovirus-Insect cells Overexpression Lysate; Human GADD45A/DDIT-1 Insect Cell Lysate (WB positive control); growth arrest and DNA-damage-inducible; alpha; Human DDIT1 Overexpression Lysate; Human GADD45 Overexpression Lysate; GADD45A cell lysate
Ordering
For Research Use Only!
Host
Baculovirus-Insect cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading buffer).
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Sequence Positions
402aa
Sequence
Met1-Arg165
Applicable Applications for GADD45A cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
Human
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.2. Re-dissolve the pellet using 200uL pure water and boil for 2-5 min.
Sequence Construction
A DNA sequence encoding the full length of human GADD45A (P24522) (Met 1-Arg 165) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for GADD45A cell lysate
GADD45A is a member of the GADD45 Family, and has been found to associate with several cytoplasmic and nuclear factors and has been implicated in several cellular functions, including MAPK signaling, cell cycle regulation, DNA repair and genomic stability, apoptosis, and immune responses. The GADD45 Family of genes is rapidly induced by different stressors, including differentiation-inducing cytokines, and there is a large body of evidence that their cognate proteins are key players in cellular stress responses. GADD45A protein has been reported to interact with multiple important cellular proteins, including Cdc2 protein kinase, proliferating cell nuclear antigen (PCNA), p21Waf1/Cip1 protein, core histone protein and MTK/MEKK4, an up-stream activator of the JNK/SAPK pathway, indicating that GADD45A may play important roles in the control of cell cycle checkpoint, DNA repair process, and signaling transduction. GADD45A expression in response to genotoxic stress illustrates a more complex scenario, wherein transcriptional changes operate in concert with mRNA turnover and translational regulation. GADD45A was the first stress-inducible gene determined to be up-regulated by p53 and is also a target for the p53 homologues, p63 and p73. The decreased GADD45A expression is also considered a survival mechanism, as cancer cells without this control can evade the apoptotic pathway leading to increased tumourigenesis. As GADD45A is an essential component of many metabolic pathways that control proliferating cancer cells, it presents itself as an emerging drug target worthy of further investigation.

This Human GADD45A overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of GADD45A protein from the overexpression lysate was verified.
Product Categories/Family for GADD45A cell lysate
References
Zhan Q. (2005) Gadd45a, a p53-and BRCA1-regulated stress protein, in cellular response to DNA damage. Mutat Res. 569(1-2): 133-43.Lal A, et al. (2006) Egad, more forms of gene regulation: the gadd45a story. Cell Cycle. 5(13): 1422-5.Hoffman B, et al. (2007) Role of gadd45 in myeloid cells in response to hematopoietic stress. Blood Cells Mol Dis. 39(3): 344-7.Rosemary Siafakas A, et al. (2009) Growth arrest and DNA damage-45 alpha (GADD45alpha). Int J Biochem Cell Biol. 41(5): 986-9.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
18,336 Da
NCBI Official Full Name
growth arrest and DNA damage-inducible protein GADD45 alpha isoform 3
NCBI Official Synonym Full Names
growth arrest and DNA-damage-inducible, alpha
NCBI Official Symbol
GADD45A
NCBI Official Synonym Symbols
DDIT1; GADD45
NCBI Protein Information
growth arrest and DNA damage-inducible protein GADD45 alpha; DDIT-1; DNA damage-inducible transcript-1; DNA-damage-inducible transcript 1; DNA damage-inducible transcript 1 protein; growth arrest and DNA-damage-inducible 45 alpha
UniProt Protein Name
Growth arrest and DNA damage-inducible protein GADD45 alpha
UniProt Gene Name
GADD45A
UniProt Synonym Gene Names
DDIT1; GADD45; DDIT-1
UniProt Entry Name
GA45A_HUMAN

NCBI Description

This gene is a member of a group of genes whose transcript levels are increased following stressful growth arrest conditions and treatment with DNA-damaging agents. The protein encoded by this gene responds to environmental stresses by mediating activation of the p38/JNK pathway via MTK1/MEKK4 kinase. The DNA damage-induced transcription of this gene is mediated by both p53-dependent and -independent mechanisms. Alternatively spliced transcript variants encoding distinct isoforms have been found for this gene.[provided by RefSeq, Dec 2010]

Uniprot Description

GADD45A: In T-cells, functions as a regulator of p38 MAPKs by inhibiting p88 phosphorylation and activity. Might affect PCNA interaction with some CDK (cell division protein kinase) complexes; stimulates DNA excision repair in vitro and inhibits entry of cells into S phase. Interacts with MAPK14. Interacts with GADD45GIP1. Interacts with PCNA. By UV irradiation, X-rays, growth arrest and alkylating agents. The induction is mediate by some kinase(s) other than PKC. Belongs to the GADD45 family. 2 isoforms of the human protein are produced by alternative splicing.

Protein type: Apoptosis; DNA repair, damage; Nuclear receptor co-regulator; Cell cycle regulation

Chromosomal Location of Human Ortholog: 1p31.2

Cellular Component: cytoplasm; nucleus

Molecular Function: protein binding

Biological Process: positive regulation of apoptosis; apoptosis; centrosome cycle; activation of MAPKKK activity; DNA damage response, signal transduction; negative regulation of protein kinase activity; positive regulation of JNK cascade; regulation of cyclin-dependent protein kinase activity; DNA repair; cell cycle arrest

Research Articles on GADD45A

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Product Notes

The GADD45A gadd45a (Catalog #AAA8114441) is a Cell Lysate produced from Baculovirus-Insect cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 402aa. AAA Biotech's GADD45A can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: Use at an assay dependent dilution. Researchers should empirically determine the suitability of the GADD45A gadd45a for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Met1-Arg16 5. It is sometimes possible for the material contained within the vial of "GADD45A, Cell Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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