Invitrogen™ AKT Multispecies In-Cell ELISA Kit, Colorimetric

In-cell ELISA Kit

Brand:  Invitrogen™ 62215

Product Code. 11538110

Description

The AKT Multispecies In-Cell ELISA Colorimetric Detection Kit is a simple method for quantifying intracellular proteins in whole cells. Principle of the method To perform the assay, cells are first plated, treated and fixed. Expression of the protein(s) of interest is monitored in wells of a microplate using target-specific primary antibodies (see the Important Product Information section for antibodies included in each kit) and a horseradish peroxidase (HRP)-conjugated detection reagent. The kit is supplied with a whole-cell stain to control for differences in cell plating, which is important when measuring relative levels of a protein with different treatments or assessing its post-translational modification (PTM) form. After staining, the results are analyzed by normalizing the absorbance (HRP activity) values to cell number, which adjusts for the cell plating differences among the wells. Rigorous validation Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

AKT (protein kinase B, PKB, RAS-alpha) is a 57kDa serine/threonine kinase that plays an important role in diverse biological responses such as regulation of metabolism, cell survival and growth by phosphorylating multiple proteins. AKT is activated by insulin, PI3K, IGF1 and various other growth and survival factors. AKT promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including forkhead transcription factors, and caspase-9. There are three mammalian isoforms of Akt: AKT1 (PKB alpha), AKT2 (PKB beta) and AKT3 (PKB gamma) with AKT2 and AKT3 being approximately 82% identical with the AKT1 isoform. Each isoform has a pleckstrin homology (PH) domain, a kinase domain and a carboxy terminal regulatory domain. AKT was originally cloned from the retrovirus AKT8, and is a key regulator of many signal transduction pathways. The AKT pathway is a major target for cancer therpeutics where AKT signaling dysfunction has implicated in many types of cancer, including cancer syndromes known as phakomatoses.

Specifications

• Accession Number: P31749, P31751, Q9Y243
• Conjugate: HRP
• Product Type: ELISA Kit
• Sufficient For: 96 microplate wells of overnight-cultured cells
• For Use With (Equipment): Colorimetric Microplate Reader
• Gene Symbol: AKT,CWS6,PKB,PKB-ALPHA,PRKBA,RAC,RAC-ALPHA,HIHGHH,PKBB,PKBBETA,PRKBB,RAC-BETA,MPPH,MPPH2,PKB-GAMMA,PKBG,PRKBG,RAC-PK-gamma,RAC-gamma,STK-2
• Label or Dye: HRP
• Regulatory Status: RUO
• Storage Requirements: 2°C to 8°C
• Test Time: 1 hr. 20 min.

• Description: Pierce AKT Colorimetric In-Cell ELISA Kit
• Immunoassay Kit Format: Multi-component kit with HRP-conjugate
• Sample Type: Supernatant
• Target Species: Human
• Gene ID (Entrez): 10000,207,208
• Kit Contents: Blocking Buffer, 20X Tris Buffered Saline, Surfact-Amps X-100 (10% Detergent), 10mL, Surfact-Amps X-100, HRP Conjugate, TMB Substrate, HRP Conjugate, Janus Green Whole-Cell Stain, TMB Stop Solution, Thin Plate Seal Assembly, Elution Buffer, Thin Plate Seal Assembly
• Quantity: 96 Tests
• Sample Volume: Supernatant, 10,000 cells
• Target: Human
• Total Assay Time: 24 hr.