- 44-504G - Invitrogen Antibodies RAF1 antibody

Submitted references 6-Formylindolo(3,2-b)Carbazole (FICZ) Modulates the Signalsome Responsible for RA-Induced Differentiation of HL-60 Myeloblastic Leukemia Cells.

Bunaciu RP, Jensen HA, MacDonald RJ, LaTocha DH, Varner JD, Yen A
PloS one 2015;10(8):e0135668

Heterogeneous effects of calorie restriction on in vivo glucose uptake and insulin signaling of individual rat skeletal muscles.

Sharma N, Sequea DA, Castorena CM, Arias EB, Qi NR, Cartee GD
PloS one 2014;8(6):e65118

Preventing the calorie restriction-induced increase in insulin-stimulated Akt2 phosphorylation eliminates calorie restriction's effect on glucose uptake in skeletal muscle.

Sharma N, Arias EB, Sequea DA, Cartee GD
Biochimica et biophysica acta 2012 Nov;1822(11):1735-40

Mechanisms for increased insulin-stimulated Akt phosphorylation and glucose uptake in fast- and slow-twitch skeletal muscles of calorie-restricted rats.

Sharma N, Arias EB, Bhat AD, Sequea DA, Ho S, Croff KK, Sajan MP, Farese RV, Cartee GD
American journal of physiology. Endocrinology and metabolism 2011 Jun;300(6):E966-78

Nitric oxide increases p21(Waf1/Cip1) expression by a cGMP-dependent pathway that includes activation of extracellular signal-regulated kinase and p70(S6k).

Gu M, Lynch J, Brecher P
The Journal of biological chemistry 2000 Apr 14;275(15):11389-96

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of c-RAF [pS621] was done on A549 cells treated with EGF (200ng/ml, 10 minutes). Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Tritonª X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with c-RAF [pS621] Rabbit Polyclonal Antibody (44504G, red histogram) or with rabbit isotype control (pink histogram) at 3-5 µg/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor¨ 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune¨ Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitates prepared from Hek293 cells overexpressing wild-type c-Raf and stimulated with EGF (1-4) or c-Raf mutant S621A (5) were resolved by SDS-PAGE on a 10% Tris-glycine gel, transferred to a PVDF membrane, and probed with a Phospho-c-Raf Ser621 antibody (Product # 44-504G).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 8 FICZ has effects beyond classical AhR transcriptional regulation. HL-60 cells were initiated in culture at 0.1 x 10 6 cells/ml with 1 muM RA, 100 nM FICZ, 1 muM alpha-NF and 1 muM beta-NF as indicated. Western blot assays of whole cell lysates are shown. Whole cell lysates were collected after 48 h. Lysates were resolved on a 12% polyacrylamide gel. 25 mug of protein was loaded in each well. Experiments were repeated at least three times. All the western blot data were quantified using ImageJ, normalized to GAPDH and then to untreated control and graphed as min to max floating bars with a line at the mean using GraphPad Prism software. P-value analysis cannot be performed on quantified blot data as this data is non-linear. A representative blot is shown for each marker.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 5 FICZ modulates RA-elicited signalsome. HL-60 cells were initiated in culture at 0.1 x 10 6 cells/ml with 1 muM RA, 100 nM FICZ, 1 muM alpha-NF and 1 muM beta-NF as indicated. Western blot assay of whole cell lysates is shown for AhR, c-Cbl, Cbl-b, CD38, Lyn, Fgr, pY416SFK, c-Raf, pS621c-Raf, pS289/296/301c-Raf, pS259c-Raf, pMEK, pERK, Vav1, Slp76. Whole cell lysates were collected after 48 h. Lysates were resolved on a 12% polyacrylamide gel. 25 mug of protein was loaded in each well. Experiments were repeated at least three times. All the western blot data were quantified using ImageJ, normalized to GAPDH and then to untreated control and graphed as min to max floating bars with a line at the mean using GraphPad Prism software. P-value analysis cannot be performed on quantified blot data as this data is non-linear. A representative blot is shown for each marker.

44-504G

Antibody data