Datasheet is currently unavailable. Try again or CONTACT US
Human ROBO1 Knockout A549 Cell Lysate
W09-K01-MT9
100 µg
Liquid (sterile filtered)
WB, SDS-PAGE
Human
Shipping info:
$50.00 to US & $70.00 to Canada for most products. Final costs are calculated at checkout.
Product Details
Human ROBO1 Knockout A549 Lysate - W09-K01-MT9
rabbit anti-ROBO1 antibody, ROBO 1, ROBO-1, hROBO-1, Roundabout homolog 1, Deleted in U twenty twenty, DUTT1, DUTT-1
Human
Target Details
ROBO1 - View All ROBO1 Products
ROBO1 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.
ROBO1 knockout A549 Clone 15 contains knockout deletions on all three copies of the ROBO1 gene in A549 cells. Each copy contains the same 23bp deletion induced by CRISPR/Cas 9. The deletion occurs in exon 2 and disrupts the sequence encoding amino acids 62 through 69, causing the downstream amino acid sequences to shift out of frame resulting in early stop codons. Validated by Sanger sequencing and Western blot.
Q9Y6N7 - UniProtKB
Application Details
SDS-PAGE, WB
Human ROBO1 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 ~181kDa is observed in ROBO1 knockout A549 when compared with unmodified A549 cell lysates by Western blot.
Cell Line Data
Human A549 (lung epithelial carcinoma)
Whole Cell Lysate
Not Stimulated
Tissue Culture
None (Control)
Formulation
2.0 mg/mL by BCA assay
1X RIPA Buffer with HALT Protease and Phosphatase Inhibitors
None
None
Shipping & Handling
Dry Ice
Store vial at -70° C or COLDER. For extended storage, aliquot contents to minimize freeze/thaw cycles.
Expiration date is one (1) year from date of receipt.
ROBO-1 (also called Roundabout homolog 1 precursor and Deleted in U twenty twenty (DUTT)) functions as a receptor for SLIT1 and SLIT2. The SLIT proteins are thought to act as a molecular guidance cue in cellular migration, including axonal navigation at the ventral midline of the neural tube and projection of axons to different regions during neuronal development. In axon growth cones, the silencing of the attractive effect of NTN1 by SLIT2 may require the formation of a ROBO1-DCC complex. ROBO-1 may also be required for lung development. ROBO-1 is a type I membrane protein. ROBO-1 is a widely expressed protein with the exception of the kidney. Defects in ROBO1 may be a cause of breast and lung cancer. ROBO-1 maps within a region of overlapping homozygous deletions characterized in both small cell lung cancer cell lines (SCLC) and in a breast cancer cell line.
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.