AGPAT2 Overexpression Lysate

Western Blot: AGPAT2 Overexpression Lysate (Adult Normal) [NBP2-08687] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for AGPAT2.

Product Discontinued

AGPAT2 Overexpression Lysate Summary

• Description: Transient overexpression lysate of 1-acylglycerol-3-phosphate O-acyltransferase 2 (lysophosphatidic acid acyltransferase, beta) (AGPAT2), transcript variant 2 The lysate was created in HEK293T cells, using Plasmid ID RC200622 and based on accession number NM_001012727. The protein contains a C-MYC/DDK Tag. You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
• Gene: AGPAT2

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
  
  Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
• Theoretical MW: 27.1 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.This product is manufactured by and sold under license from OriGene Technologies and its use is limited solely for research purposes.

Alternate Names for AGPAT2 Overexpression Lysate

• 1-acylglycerol-3-phosphate O-acyltransferase 2 (lysophosphatidic acidacyltransferase, beta)
• 1-acylglycerol-3-phosphate O-acyltransferase 2
• 1-AGPAT 2,1-acylglycerol-3-phosphate O-acyltransferase 2 (lysophosphatidic acidacyltransferase-beta)
• 1-AGPAT2,1-AGP acyltransferase 2,1-acyl-sn-glycerol-3-phosphate acyltransferase beta
• Berardinelli-Seip congenital lipodystrophy
• BSCL
• BSCL1
• EC 2.3.1.51
• LPAAB
• LPAAT-beta
• Lysophosphatidic acid acyltransferase beta
• lysophosphatidic acid acyltransferase-beta

Background

AGPAT2 encodes a member of the 1-acylglycerol-3-phosphate O-acyltransferase family. The protein is located within the endoplasmic reticulum membrane and converts lysophosphatidic acid to phosphatidic acid, the second step in de novo phospholipid biosynthesis. Mutations in this gene have been associated with congenital generalized lipodystrophy (CGL), or Berardinelli-Seip syndrome, a disease characterized by a near absence of adipose tissue and severe insulin resistance. Alternate transcriptional splice variants, encoding different isoforms, have been characterized. [provided by RefSeq]

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Bioinformatics

• Gene Symbol: AGPAT2