Application Note: Human MAPK1 (ERK2) Knockout A549 Cell Lysate has been tested by SDS-PAGE and western blot and is suitable for use in Western blot, ELISA, Immunoprecipitation and ChIP. No detection of expected band at ~44kDa is observed in MAPK1 (ERK2) knockout A549 when compared with unmodified A549 cell lysates by Western blot.
Cell Line: Human A549 (lung epithelial carcinoma)
Clone ID: Clone 15
Concentration Value: 2.0 mg/mL
ELISA Dilution: User Optimized
ChIP Dilution: User Optimized
Immunoprecipitation Dilution: User Optimized
Western Blot Dilution: User Optimized
General Disclaimer Note: This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
Physical State: Liquid (sterile filtered)
Purity and Specificity: MAPK1 (ERK2) knockout A549 cells were grown in Dulbecco’s medium supplemented with 10% fetal bovine serum. Cells were washed with PBS and then incubated on ice in modified RIPA buffer to lyse the cells. Protein integrity was ensured using a cocktail of protease inhibitors with broad specificity for the inhibition of aspartic, cysteine, and serine proteases as well as aminopeptidases (0.1 mM AEBSF HCl, 0.08 µM Aprotinin, 5 µM Bestatin, 1.5 µM E-64, 2 µM Leupeptin Hemisulfate, 1 µM Pepstatin A). Phosphatase inhibitors 1 mM NaF and 1 mM Na3VO4 were also added. Cell debris was removed by centrifugation. Protein concentration was determined by BCA using a commercially available kit. Protein concentration was adjusted to 2 mg/ml with modified 1X RIPA buffer.MAPK1 (ERK2) knockout A549 Clone 15 contains knockout deletions on all three copies of the MAPK1 (ERK2) gene in A549 cells. Each copy contains the same 104bp deletion induced by CRISPR/Cas9. The deletion occurs in exon 2 and disrupts the sequence between amino acids 59 to 94. These mutations induce a frame-shift and result in early stop codons. Validated by Sanger sequencing and Western blot.
Background: ERK2 antibodies detect the ERK2 isoform. Mitogen activated protein kinase 1, also known as MAPK1, ERK, or ERK2, is an integral component of the MAP kinase cascade that regulates cell growth and differentiation. ERK1 and ERK2 are activated by MEK1 and MEK2 in the B-raf signaling pathway resulting in its translocation to the nucleus where it phosphorylates nuclear targets. Human ERK1 and ERK2 are 84% identical in sequence and share common functionality in cells.
Low Endotoxin: No
English
Deutsch
