Cytoskeleton now offers the first and only commercially available RhoA ELISA.
It is recommended to use the BK150 kit in conjunction with the RhoA G-LISA™ kit (BK124), allowing quantitation of Total RhoA and Active RhoA in the same lysates.
Providing precise solutions to RhoA quantification, enabling researchers to...
Kit Contents - Enough reagents for 96 assays.
Lachowski, D. et al. G Protein-Coupled Estrogen Receptor Regulates Actin Cytoskeleton Dynamics to Impair Cell Polarization. Front. Cell Dev. Biol. 8, 1127 (2020).
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Ngai, D. et al. DDR1 (Discoidin Domain Receptor-1)-RhoA (Ras Homolog Family Member A) Axis Senses Matrix Stiffness to Promote Vascular Calcification. Arterioscler. Thromb. Vasc. Biol. 40, 1763–1776 (2020).
Dér, B. et al. NK2 receptor-mediated detrusor muscle contraction involves G q/11 -dependent activation of voltage-dependent Ca 2+ channels and the RhoA-Rho kinase pathway. Am. J. Physiol. Physiol. 317, F1154–F1163 (2019).
Giannini, M. et al. Nano-topography: Quicksand for cell cycle progression? Nanomedicine Nanotechnology, Biol. Med. 14, 2656–2665 (2018).
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M.J. Herr et al., 2014. Tetraspanin CD9 regulates cell contraction and actin arrangement via RhoA in human vascular smooth muscle cells. PLoS ONE. 9:e106999.
Valtcheva et al., 2013. The orphan adhesion G protein coupled receptor GPR97 regulates migration of lymphatic endothelial cells via the small GTPases RhoA and Cdc42. J. Biol. Chem. doi: 10.1074/jbc.M113.512954.
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Question 1: Can I use the same lysate samples to measure total RhoA with this kit (Cat. # BK150) and activated RhoA samples in the G-LISA activation assay kit (Cat. # BK124)?
Answer 1: Yes, the same lysate samples can be used in both the total RhoA ELISA (Cat. # BK150) and RhoA G-LISA activation assay kit (Cat. # BK124). When preparing samples to be used with both kits, please use the lysis buffer (Part # GL36) that comes with the RhoA G-LISA assay kit. Generally G‐LISA samples have low protein concentrations, e.g., 0.3 to 0.6 mg/ml. At this concentration the samples are at the lower range of detection for the ELISA. Therefore we recommend using more lysate and less Sample Dilution Buffer when preparing G-LISA samples for the ELISA. To keep the final concentration higher, we recommend using 40 μl of G‐LISA extract plus 80 μl of Sample Dilution Buffer, which is sufficient for duplicate ELISA wells. In this case, also use the same Lysis:SDB ratio for the positive control protein solutions.
Question 2: I see that this ELISA uses a 96 well plate format. Do I have to use all 96 wells at one time or can I save the unused wells for use at a later date?
Answer 2: The 96 well plates are packaged as 12 x 8 well strips. Thus, each strip is removable and can be further broken down to a 1 well format for an assay. In this way, there is tremendous format flexibility since 1-96 wells can be used in an assay. It is imperative to keep the plate in the sealed desiccant bag with desiccant at all times. Move to room temperature 30 min prior to starting the assay. Open the bag and remove the number of strips or wells needed immediately prior to the start of the experiment, place the remaining strips/wells in the desiccated bag, reseal and return to storage at 4°C. Prepare and use the strips/wells as directed.
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