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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [4]
- Immunocytochemistry [1]
- Other assay [1]
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- Product number
- PA5-29931 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Rhotekin Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, Jurkat, Raji, mouse brain, NCI-H929, human Rhotekin-transfected 293T.
- Concentration
- 0.54 mg/mL
Submitted references S100A4 is activated by RhoA and catalyses the polymerization of non-muscle myosin, adhesion complex assembly and contraction in airway smooth muscle.
Zhang W, Gunst SJ
The Journal of physiology 2020 Oct;598(20):4573-4590
The Journal of physiology 2020 Oct;598(20):4573-4590
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Supportive validation
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using Rhotekin Polyclonal Antibody (Product # PA5-29931). Sample (30 µg of whole cell lysate). Lane A: JurKat . 7.5% SDS PAGE. Rhotekin Polyclonal Antibody . Rhotekin Polyclonal Antibody (Product # PA5-29931) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using Rhotekin Polyclonal Antibody (Product # PA5-29931). Sample (20 µg of whole cell lysate). Lane A: mouse brain . 7.5% SDS PAGE. Rhotekin Polyclonal Antibody (Product # PA5-29931) diluted at 1:10,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Rhotekin Polyclonal Antibody detects Rhotekin protein by western blot analysis. A. 30 µg 293T whole cell lysate/extract. B. 30 µg whole cell lysate/extract of human Rhotekin-transfected 293T cells.7.5 % SDS-PAGE. Rhotekin Polyclonal Antibody (Product # PA5-29931) dilution: 1:5,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Rhotekin Polyclonal Antibody detects Rhotekin protein by western blot analysis. A. 30 µg Jurkat whole cell lysate/extract. B. 30 µg Raji whole cell lysate/extract. C. 30 µg NCI-H929 whole cell lysate/extract.7.5 % SDS-PAGE. Rhotekin Polyclonal Antibody (Product # PA5-29931) dilution: 1:5,000.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Rhotekin showing staining in the cytoplasm of HeLa cells. Rhotekin-transfected (right) or untransfected (left) HeLa cells were fixed in ice-cold MeOH for 5 min and stained using a Rhotekin polyclonal antibody (Product # PA5-29931) diluted at 1:500. Blue: Hoechst 33342 staining.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- 5 Figure Rhotekin couples RhoA to S100A4 and NM myosin II in SM cells and tissues stimulated with ACh A , rhotekin (RTKN) was immunoprecipitated from tissue extracts and immunocomplexes were blotted for RhoA, S100A4, RTKN and NM myosin IIA. Stimulation for 5 min with 10 -5 M ACh significantly increased the coprecipitation of NM myosin IIA ( n = 6, p = 0.0070), RhoA ( n = 3, p = 0.0359) and S100A4 ( n = 9, p = 0.0001) with RTKN. B , in situ PLA was used to determine the interaction of RTKN and RhoA, S100A4 and NM myosin IIA in freshly dissociated tracheal SM cells. Stimulation with ACh caused a significant increase in the number of PLA complexes between all four proteins (RTKN-RhoA, n = 20, p = 0.0001; RTKN-S100A4, n = 20, p = 0.0001; RTKN-NM myosin IIA, n = 31, p = 0.0001). C , tracheal SM tissues were treated with RTKN siRNA. Depletion of RTKN significantly inhibited protein expression ( n = 10, p = 0.0001) and also inhibited tension development in response to 10 -5 M ACh stimulation ( n = 16, p = 0.0001). D , PLA shows interactions between S100A4 and RhoA in cells dissociated from sham-treated tissues and from RTKN-depleted tissues. ACh stimulation of sham-treated cells resulted in a significant increase in the interactions between S100A4 and RhoA (US, n = 16; ACh, n = 20). RTKN depletion inhibited the ACh-induced increase of the interaction between S100A4 and RhoA ( p = 0.0001) (US, n = 18; ACh, n = 19). Data analysed by a paired Student's t test ( A and C ), an unpaired S
