Ab729

Manufactured by Abcam

Sourced in United States

Ab729 is a laboratory product manufactured by Abcam. It is a general-purpose piece of lab equipment. The core function of Ab729 is to facilitate standard laboratory procedures. No further details are available at this time.

Automatically generated - may contain errors

Lab products found in correlation

Nb100 449, by Novus Biologicals (1 mentions) Image lab software, by Bio-Rad (1 mentions) Nbp1 19751, by Novus Biologicals (1 mentions) Ab18061, by Abcam (1 mentions) Mab377, by Merck Group (1 mentions) T5168, by Abcam (1 mentions) Ab15452, by Merck Group (1 mentions) Mab3420, by Merck Group (1 mentions) Ab6881, by Abcam (1 mentions) Fitc conjugated donkey anti goat igg, by Abcam (1 mentions) Triton x 100, by Merck Group (1 mentions) Fish gelatin, by Merck Group (1 mentions) Lsm 700 confocal microscope, by Zeiss (1 mentions) Alexa fluor 488 conjugated anti mouse, by Thermo Fisher Scientific (1 mentions)

4 protocols using ab729

Western Blot Analysis of Angiogenic Factors

Check if the same lab product or an alternative is used in the 5 most similar protocols

Treated cells were lysed using radioimmunoprecipitation assay (RIPA) lysis buffer for whole-cell lysates, containing protease and phosphatase inhibitors. Protein concentrations were determined using the Pierce Bicinchoninic (BCA) Protein Assay Kit, and total protein concentration between 10 and 20 μg of protein was loaded for Western blot analysis. Membranes were probed with antibodies for BMP2 (NBP119751, Novus Biologicals, Centennial, CO, USA, 1:1000), total HIF-1α (NB100–449, Novus Biologicals, 1:500), VEGFA (ab461154, Abcam, Trumpington, Cambridge, UK, 1:2000), total VEGFR2 (2479; Cell Signalling Technology, Danvers, MA, USA, 1:1000), phosphorylated (y¹¹⁷⁵) VEGFR2 (2478, Cell Signalling Technology, 1:1000), total eNOS (610297, BD Biosciences, San Jose, CA, USA, 1:1000), and phosphorylated (S¹¹⁷⁷) eNOS (61293, BD Biosciences, 1:1000). Even loading was confirmed by α-tubulin (ab729, Abcam, 1:5000) for total lysate. Protein levels were quantified and analysed using Bio-Rad ImageLab software (v.5.0, 170-9692, Bio-Rad Laboratories, Gladesville, CA, USA).

Mulangala J., Akers E.J., Solly E.L., Bamhare P.M., Wilsdon L.A., Wong N.K., Tan J.T., Bursill C.A., Nicholls S.J, & Di Bartolo B.A. (2022). Pro-Calcific Environment Impairs Ischaemia-Driven Angiogenesis. International Journal of Molecular Sciences, 23(6), 3363.

+ Open protocol

+ Expand

Immunolabeling of Neuronal Cytoskeleton

Check if the same lab product or an alternative is used in the 5 most similar protocols

Neurons were fixed with pre-warmed 4 % paraformaldehyde/4 % sucrose in PBS for 15 min, permeabilized with 0.1 % Triton X-100 in PBS for 10 min, and blocked with a blocker solution (2 % bovine serum albumin, 2 % fetal bovine serum, and 0.2 % fish gelatin in PBS) for 1 h at room temperature, as described previously [4 (link)]. Cells were then incubated the primary antibodies, including a mouse monoclonal anti-α-tubulin antibody (1:1000, Sigma, no. T-5168), a goat polycolonal anti-TRPM7 (1:200, Abcam, Ab729), a mouse monoclonal anti-α actinin (1:500, Abcam, Ab18061), or a mouse monoclonal anti-NeuN (1:500, Millipore, no. MAB377). To differentiate between axons and dendrites, neurons were incubated with mouse monoclonal anti-tau-1 (axonal marker; 1:500, Millipore, MAB3420) and chicken polyclonal anti-microtubule associated protein 2 (MAP2) (dendritic marker; 1:500, Millipore, Ab15452).

Turlova E., Bae C.Y., Deurloo M., Chen W., Barszczyk A., Horgen F.D., Fleig A., Feng Z.P, & Sun H.S. (2014). TRPM7 Regulates Axonal Outgrowth and Maturation of Primary Hippocampal Neurons. Molecular neurobiology, 53(1), 595-610.

+ Open protocol

+ Expand

TRPM7 Localization in HEK293 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Stable HEK293 cells were plated onto glass coverslips (⌀13 mm) that were coated with Matrigel, and protein expression was induced by doxycycline (1 µg/ml) for 24 h. Cells were fixed in 4% formaldehyde for 10 min at room temperature (RT), washed three times with PBS, and permeabilized in 0.2% Triton X-100 (Sigma-Aldrich) in PBS for 10 min at RT. Cells were washed three times with cold PBS and incubated with 1% BSA in PBS for 10 min and then with goat anti-TRPM7 antibody (ab729; Abcam) for 16 h at 4°C. After three washes with 1% BSA-PBS, cells were incubated with a secondary antibody, FITC-conjugated donkey antigoat IgG (ab6881; Abcam). After three washes with 1% BSA-PBS, coverslips were mounted for confocal microscopic imaging (FluoView version 10; Olympus).

Inoue H., Murayama T., Kobayashi T., Konishi M, & Yokoyama U. (2021). The zinc-binding motif of TRPM7 acts as an oxidative stress sensor to regulate its channel activity. The Journal of General Physiology, 153(6), e202012708.

+ Open protocol

+ Expand

Imaging TRPM7 Localization in U87 and NHA Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

U87 and NHA cells (5×10⁴ cells/mL) were maintained on 18-mm round Poly-D-Lysine-coated coverslips (Warner Instruments, USA) for 24 hours, fixed with 4% paraformaldehyde for 20 minutes at RT, and then permeabilized for 20 min with 0.1% Triton X-100 in PBS. Cells were incubated overnight at 4°C with anti-TRPM7 (ab729, Abcam, 1:50) and β-tubulin (mouse mAb, Invitrogen, USA, 1:500) antibodies in 2% bovine serum albumin (BSA, Bioshop, Canada), 2% FBS and 0.2% fish gelatin (Sigma-Aldrich, USA). The cells were then incubated with Alexa-Fluor 488 conjugated anti-mouse and Alexa-Fluor 405 conjugated anti-goat (1:500, Molecular Probes, USA) antibodies for 2 hours at RT. Fluorescence was visualized with an LSM700 Zeiss confocal microscope (Carl Zeiss Inc., Gottingen, Germany).

Chen W.L., Barszczyk A., Turlova E., Deurloo M., Liu B., Yang B.B., Rutka J.T., Feng Z.P, & Sun H.S. (2015). Inhibition of TRPM7 by carvacrol suppresses glioblastoma cell proliferation, migration and invasion. Oncotarget, 6(18), 16321-16340.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!