44-496G

Lebron MB, Brennan L, Damoci CB, Prewett MC, O'Mahony M, Duignan IJ, Credille KM, DeLigio JT, Starodubtseva M, Amatulli M, Zhang Y, Schwartz KD, Burtrum D, Balderes P, Persaud K, Surguladze D, Loizos N, Paz K, Kotanides H
Cancer biology & therapy 2014 Sep;15(9):1208-18

Lyn contributes to regulation of multiple Kit-dependent signaling pathways in murine bone marrow mast cells.

Shivakrupa R, Linnekin D
Cellular signalling 2005 Jan;17(1):103-9

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Peptide Competition: Extracts prepared from M07e cells left untreated (1) or treated (2-5) with SCF were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then we

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of C-Kit (pY730) was performed by loading 30 µg of A431 (lane1), A431 treated for 10 minutes with 200 ng/mL of EGF (lane2), Rat Lung (lane3), HeLa (lane4), A549 (lane5) and Serum Starved A549 (lane6) lysate using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. C-Kit (pY730) was detected at ~170 kDa using C-Kit (pY730) Rabbit Polyclonal Antibody (Product # 44-496G) at 1:1000 dilution in 5 % skim milk at 4ºC overnight on a rocking platform. Goat Anti-Rabbit IgG - HRP Secondary Antibody (G21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of Phospho c-kit (Tyr 730), showing staining in the cytoplasm and membrane of paraffin-embedded human gastric carcinoma tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Phospho-c-Kit (Tyr730), Polyclonal Antibody (Product # 44-496G) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL