Anti alk5

Manufactured by Abcam

Sourced in United Kingdom

Anti-ALK5 is a laboratory reagent that targets the ALK5 protein. It is used for research purposes in the scientific community.

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Lab products found in correlation

Anti gapdh, by Abcam (1 mentions) Anti smad2 3, by Santa Cruz Biotechnology (1 mentions) Anti p smad2 3, by Santa Cruz Biotechnology (1 mentions) Anti gapdh, by Merck Group (1 mentions) Horseradish peroxidase linked secondary antibody, by Santa Cruz Biotechnology (1 mentions) Any kd mini protean tgx precast protein gel, by Bio-Rad (1 mentions) Polyvinylidene fluoride membrane, by Bio-Rad (1 mentions) Ripa lysis buffer, by Merck Group (1 mentions) Supersignal west pico plus chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Rabbit anti-ALK5 (3 citations)

4 protocols using anti alk5

Quantitative Western Blot Analysis

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Total cellular proteins were obtained by RIPA lysis buffer with protease and phosphatase inhibitors. The protein concentration was calculated by the BCA protein assay kit (Pierce), and 35 μg of protein lysate was loaded onto a 12% SDS-PAGE gel. After electrophoresis, the proteins were transferred to a hydrophobic PVDF membrane (Millipore), which was then blocked by incubation with 5% non-fat dry milk at room temperature for 1 h. The membranes were incubated with the primary antibodies anti-ALK5 (1:1000; Abcam, Cat# ab31013) and anti-GAPDH (Abcam, Cat# ab9485) at 4 °C overnight and washed three times with 1× TBST. After incubation with secondary antibodies, signals were visualised by a chemiluminescence detection system. GAPDH was used as the loading control.

Shi C.J., Lian J.J., Zhang B.W., Cha J.X., Hua Q.H., Pi X.P., Hou Y.J., Xie X, & Zhang R. (2022). TGFβR-1/ALK5 inhibitor RepSox induces enteric glia-to-neuron transition and influences gastrointestinal mobility in adult mice. Acta Pharmacologica Sinica, 44(1), 92-104.

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Peri-infarct Area Protein Analysis

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Rats were anesthetized and decapitated on a given day, and brain tissues from the ipsilateral peri-infarct area were collected and extracted. Anti-ALK5 (1:1000, Abcam), anti-Smad2/3 (1:1000; Santa Cruz) and anti-pSmad2/3 (1:1000; Santa Cruz) were used as primary antibodies. After the secondary antibody reaction, the bands were visualized by electrochemiluminescence (Millipore, Darmstadt, Germany). The positive pixel area was detected by an image analysis software (BIO-RAD Gel Doc 2000, Watertown, MA, USA). Western blot quantification was performed by densitometry and was normalized to GAPDH. All experiments were conducted under the same conditions and repeated three times.

Liu Z., Liu M., Jia G., Li J., Niu L., Zhang H., Qi Y., Sun H., Yan L.J, & Ma J. (2023). Long-term intermittent fasting improves neurological function by promoting angiogenesis after cerebral ischemia via growth differentiation factor 11 signaling activation. PLOS ONE, 18(3), e0282338.

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Western Blot Protein Analysis Protocol

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Protein extraction was performed using a previously described protocol [80 (link)]. Lysates were resolved by sodium dodecyl sulfate polyacrylamide gel electrophoresis, using a 12% acrylamide gel. Proteins were then transferred to nitrocellulose membranes, and subsequently probed overnight with the following antibodies: anti-Alk5 (Abcam, Cambridge, UK) at 1:1000 and anti-GAPDH (Millipore, Watford, UK) at 1:30000.

Hodgson D., Rowan A.D., Falciani F, & Proctor C.J. (2019). Systems biology reveals how altered TGFβ signalling with age reduces protection against pro-inflammatory stimuli. PLoS Computational Biology, 15(1), e1006685.

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Immunoblotting Analysis of Protein Signaling

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Protein was extracted by RIPA lysis buffer (Sigma) and resolved in Any kD™ Mini-PROTEAN® TGX™ Precast Protein Gels (Bio-Rad), transferred to a polyvinylidene fluoride membrane (Bio-Rad) and blocked with 5% BSA at room temprature for 1 hour and analyzed by immuno-blotting with the following antibodies: anti-SKI (G8, Santa Cruz), anti-ALK4 (EPR4815, Abcam), anti-ALK5 (Abcam), anti-p-Smad2 (S465/467) (138D4, CST), anti-Smad2 (D43B4, CST), anti-p-ERK1/2 (Thr202/Tyr204) (CST), anti-ERK1/2 (clone 16, BD), anti-p-STAT3 (Tyr705) (3E2, CST), anti-STAT3 (124H6, CST), and anti-β-actin antibodies (I-19, Santa Cruz), at 4°C overnight. The blot was subsequently incubated with horseradish peroxidase-linked secondary antibodies (Santa Cruz) at room temprature for 1 hour and the signal was detected using SuperSignal™ West Pico PLUS Chemilum inescent Substrate (Thermofisher). The relative protein amounts of SKI, ALK4, ALK5 to endogenous control β-actin, p-Smad2 to Smad2, and p-ERK1/2 to ERK1/2 were quantified by densitometric analysis using Image J software (Schneider et al., 2012 (link)) and presented using arbitrary units.

Wu B., Zhang S., Guo Z., Bi Y., Zhou M., Li P., Seyedsadr M., Xu X., Li J.L., Markovic-Plese S, & Wan Y.Y. (2021). The TGF-β superfamily cytokine Activin-A is induced during autoimmune neuroinflammation and drives pathogenic Th17 cell differentiation. Immunity, 54(2), 308-323.e6.

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