PA5-29931

antibody from Invitrogen Antibodies

Targeting: RTKN B5

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-29931

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. Predicted reactivity: Mouse (93%), Rat (96%), Rhesus Monkey (99%), Bovine (94%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.54 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

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

Immunocytochemistry

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.

Other assay

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