PA5-29767

antibody from Invitrogen Antibodies

Targeting: TAGLN DKFZp686P11128, SM22, SMCC, TAGLN1, WS3-10

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-29767

Provider

Invitrogen Antibodies

Product name

TAGLN Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: 293T, A431, HeLa, HepG2, Mouse liver, A-10, A-10 treated with MG132, Bovine Smooth Muscle Cell, Bovine Fibroblastic Chondrocyte. Predicted reactivity: Mouse (97%), Rat (96%), Xenopus laevis (81%), Pig (98%), Chicken (87%), Bovine (97%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse, Rat, Bovine

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

0.33 mg/mL

Storage

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

References

Submitted references

Transgelin is a poor prognostic factor associated with advanced colorectal cancer (CRC) stage promoting tumor growth and migration in a TGFβ-dependent manner.

Elsafadi M, Manikandan M, Almalki S, Mahmood A, Shinwari T, Vishnubalaji R, Mobarak M, Alfayez M, Aldahmash A, Kassem M, Alajez NM
Cell death & disease 2020 May 11;11(5):341

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Transgelin is a TGFβ-inducible gene that regulates osteoblastic and adipogenic differentiation of human skeletal stem cells through actin cytoskeleston organization.

Elsafadi M, Manikandan M, Dawud RA, Alajez NM, Hamam R, Alfayez M, Kassem M, Aldahmash A, Mahmood A
Cell death & disease 2016 Aug 4;7(8):e2321

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemistry-Immunofluorescence analysis of TAGLN was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: TAGLN Polyclonal Antibody (Product # PA5-29767) diluted at 1:100. Red: alpha Tubulin, a cytoskeleton marker. Blue: Hoechst 33342 staining.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 3 TAGLN overexpression induces osteogenesis and adipogenesis in hMSC. ( a ) All controls represent empty vector-transfected cells. qRT-PCR of TAGLN gene expression in control (empty vector) hMSC and TAGLN-overexpressing line (TAGLN-hMSC). ( b ) Western blotting for TAGLN in TAGLN-hMSC (upper panel) and B-Actin (lower panel). ( c ) TAGLN immunocytochemical staining in control hMSC and TAGLN-hMSC cells at magnification x 40 and x 60. Cells were differentiated into osteoblasts by osteogenic or adipogenic induction mixture for 14 days. ( d ) hMSC and TAGLN-hMSC cells were implanted with HA/TCP subcutaneously into NOD/SCID mice. The histology of in vivo formed bone was examined with H&E staining (original magnification x 200). High-resolution whole-slide digital scans of all histological sections were created with a ScanScope scanner and bone formed was quantified. ( e ) Mineralized bone matrix formation in control hMSC (empty vector) cells and TAGLN-hMSC stained for Alizarin Red. ( f ) Quantification of Alizarin Red staining in control hMSC and TAGLN-hMSC. ( g ) qRT-PCR for osteogenic markers: ALPL, OSC, COL1A1 (collagen type I), and OSP: NI, OS, and osteoblast-induced TAGLN-hMSC (OS TAGLN-hMSC). ( h ) Both control (empty vector) hMSC, and TAGLN-hMSC were induced to adipocyte differentiation using adipocyte induction medium for 7 days. Mature lipid-filled adipocytes were stained by oil red. ( i ) Quantification of Nile Red staining for mature adipocytes: control (empty vec

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 4 Effects of TAGLN overexpression on hFFs functions. ( a ) qRT-PCR of TAGLN gene expression in control (empty vector) TAGLN-hFFs. ( b ) Immunocytochemical staining for TAGLN protein in control hFFs, and TAGLN-hFFs at magnifications x 40 and x 60. ( c ) Western blot of TAGLN (upper panel) and B-Actin (lower panel) in control hFFs and TAGLN-hFFs. Cells were induced to differentiate into osteoblasts or adipocytes by either osteogenic or adipogenic mixture for 24 days. ( d ) Alamar blue staining for cell viability in control hFFs and TAGLN-hFFs on day (D): 1D, 2D, 3D, and 4D. ( e ) Scratch assay where the motility of hFFs and TAGLN-hFFs cells was observed at different time intervals: 0, 24, 30, and 50 h. ( f ) Alizarin Red S staining for mineralized matrix formation. ( g ) Quantification of mineralized matrix formation in control hFFs cells and TAGLN-hFFs using Osteo-Image Mineralization Assay. ( h ) Control hFFs and hFF-TAGLN were induced to adipocyte differentiation by incubation in adipocyte induction medium for 24 days and stained by oil red for lipid-filled mature adipocytes. ( i ) Quantification of Nile red staining of mature adipocytes formed in control hFF cells and TAGLN-hFFs. Data are shown as mean+-S.D. of three independent experiments. ** P

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 2 TAGLN is a TGFbeta responsive gene in CRC. a TAGLN mRNA expression in three different CRC cell lines based on the CCLE database: HCT116, HT-29, and RKO. Expression level is compared to TAGLN expression in HCT116 cells. b qRT-PCR of TAGLN expression in HCT116 control cells or cells transduced with TAGLN-overexpressing lentiviral vector (upper panel). Western blotting for TAGLN (upper panel) in HCT116-TAGLN vs. control cells. beta-Actin was used as the loading control. QRT-PCR for TAGLN ( c ) or TPM1 ( d ) gene expression 3 days post transfection of RKO cells with TAGLN-siRNA (siTAGLN) or scrambled-siRNA (siScr). Data are presented as fold change in mRNA expresion. e qRT-PCR of TAGLN, TPM1, and ACTA2 gene expression in RKO cells treated with TGFbeta1 (10 ng/ul) or SB431542 (10 µM), compared to control cells. The two-tailed t -test was used to compare different treatment groups. ** P < 0.005, *** P < 0.0005.