Invitrogen

SCXA Polyclonal Antibody

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SCXA Antibody in Western Blot (WB) — Western blot analysis using a SCXA polyclonal antibody (Product # PA5-23943) in mouse cerebellum tissue lysates (35 µg per lane).

SCXA Antibody — Figure 5 The origin of cells affects the formation of proteins associated with tenogenesis. Left panel: TGF- beta 2 secretion determined by ELISA. Results shown as the concentration in the supernatant per 10 * 10 3 cells. Right panel: SMAD3, COL14A1, MKX, and SCX determined by Western blot. Expression of beta -actin was used as a loading control. The results shown as integrated optical density (IOD) normalized to IOD of corresponding beta -actin. Data presented as mean +- SEM. Statistical analysis was performed by Student's t -test ( t ) or Mann-Whitney U test (U). p < 0.05 was assumed to be statistically significant and highlighted in red. TGF- beta 2: TRANSFORMING GROWTH FACTOR BETA 2; SMAD3: SMAD FAMILY MEMBER 3; COL14A1: COLLAGEN TYPE XIV ALPHA 1; MKX: MOHAWK HOMEOBOX; SCX: SCLERAXIS.

SCXA Antibody — Figure 3 cANGPTL4 regulates COL1A2 and COL3A1 expression in fibroblasts via a bHLH-dependent signaling mechanism. ( a ) Immunoblot analysis of indicated proteins from fibroblasts treated with cANGPTL4 (12 ug/mL) in the presence or absence of TGFbeta1 (10 ng/mL). beta-tubulin, as loading control, was from the same samples. ( b,d ) Chromatin immunoprecipitation (ChIP) of SBE sites in the human COL1A2 gene promoter region and E-box sites in the human COL1A2 and COL3A1 gene promoter regions using phospho-Smad3 antibodies and scleraxis (ScxA) antibodies respectively. The gene sequences spanning the SBE and E-box sites and a random control sequence were analyzed by PCR from immunoprecipitated chromatin from fibroblasts treated with TGFbeta1 in the presence or absence of cANGPTL4 (cANG). Pre-immune serum was used as a control. PCR was performed using chromatin before immunoprecipitation as input. M, 100 bp DNA ladder. ( c ) A schematic diagram showing the relative positions of Enhancer boxes (E-Boxes) and Smad binding elements (SBE) on the human COL1A2 and COL3A1 proximal promoters. Sequences of E-Boxes and SBE are shown. ChIP PCR primers designed specifically to bind scleraxis along this proximal promoter region are indicated. ( e ) Relative mRNA expression of ID3 in fibroblasts treated with 12 ug/mL of cANGPTL4 and lysed for mRNA at the indicated time points post-treatment. ( f ) Relative mRNA expression of ID3 in control (F ctrl ) and ID3-knockdown (F ID3 ) fibroblasts treated fo

SCXA Antibody — Figure 3 cANGPTL4 regulates COL1A2 and COL3A1 expression in fibroblasts via a bHLH-dependent signaling mechanism. ( a ) Immunoblot analysis of indicated proteins from fibroblasts treated with cANGPTL4 (12 ug/mL) in the presence or absence of TGFbeta1 (10 ng/mL). beta-tubulin, as loading control, was from the same samples. ( b,d ) Chromatin immunoprecipitation (ChIP) of SBE sites in the human COL1A2 gene promoter region and E-box sites in the human COL1A2 and COL3A1 gene promoter regions using phospho-Smad3 antibodies and scleraxis (ScxA) antibodies respectively. The gene sequences spanning the SBE and E-box sites and a random control sequence were analyzed by PCR from immunoprecipitated chromatin from fibroblasts treated with TGFbeta1 in the presence or absence of cANGPTL4 (cANG). Pre-immune serum was used as a control. PCR was performed using chromatin before immunoprecipitation as input. M, 100 bp DNA ladder. ( c ) A schematic diagram showing the relative positions of Enhancer boxes (E-Boxes) and Smad binding elements (SBE) on the human COL1A2 and COL3A1 proximal promoters. Sequences of E-Boxes and SBE are shown. ChIP PCR primers designed specifically to bind scleraxis along this proximal promoter region are indicated. ( e ) Relative mRNA expression of ID3 in fibroblasts treated with 12 ug/mL of cANGPTL4 and lysed for mRNA at the indicated time points post-treatment. ( f ) Relative mRNA expression of ID3 in control (F ctrl ) and ID3-knockdown (F ID3 ) fibroblasts treated fo

SCXA Antibody — Figure 5 cANGPTL4 upregulates ID3 via a beta-catenin-dependent pathway. ( a ) Immunoblot of full-length cadherin-11 (CDH11) and splice variant CDH11 (CDH11 var ) in human fibroblasts. Fibroblasts were transfected with empty vector (F ctrl ) or expression vector containing cDNA encoding CDH11 var (F CDH11var ). beta-tubulin, as loading control, was from the same samples. Relative mRNA expression of ID3 in fibroblasts subjected to the indicated treatments. ( b-e ) Immunodetection and quantification of ID3 or beta-catenin proteins in fibroblasts subjected to the indicated treatments. beta-tubulin (cytosol) and c-JUN (nuclear) from the same samples were used as loading controls. Immunoblot images are cropped; full-length immunoblots can be found in Supplementary Fig. S4h-n . ( f , g ) Relative mRNA and protein expression of ID3 in fibroblasts treated with ICG001 and XAV939, two inhibitors of beta-catenin downstream signaling. Ribosomal protein L27 was used as housekeeping control for qPCR. beta-tubulin from the same samples was used as loading control for immunoblots. Values represent mean +- SD, n = 5 independent experiments. n.s., not significant, *P < 0.05, **P < 0.01, ***P < 0.001 (Mann-Whitney U-test). ( h ) ChIP of TBE on the human ID3 gene using anti-beta-catenin antibody followed by re-ChIP with anti-LEF antibody. Gene sequence spanning the TBE II/III and a random control sequence were analyzed by PCR in the immunoprecipitated chromatin of fibroblasts subjected to the ind

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1:1000

View 2 publications 2 publications

ChIP assay (ChIP)

-

View 1 publication 1 publication

Product Specifications

Species

Human, Mouse

Published species

Not Applicable

Expression System

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

KLH conjugated synthetic peptide between 172-201 amino acids from the C-terminal region of human SCXA

Conjugate

Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Protein A, Antigen affinity chromatography

Storage buffer

PBS

Contains

0.09% sodium azide

Storage conditions

-20° C, Avoid Freeze/Thaw Cycles

RRID

AB_2541443

Product Specific Information

This antibody is predicted to react with chicken based on sequence homology.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: Basic helix-loop-helix transcription factor scleraxis; bHLHa41; bHLHa48; Class A basic helix-loop-helix protein 41; Class A basic helix-loop-helix protein 48; class II bHLH protein scleraxis; scleraxis basic helix-loop-helix transcription factor; scleraxis homolog A; scleraxis homolog B

Gene Aliases: BB114693; BHLHA41; BHLHA48; Scl; SCX; SCXA; SCXB

UniProt ID: (Human) Q7RTU7, (Mouse) Q64124

Entrez Gene ID: (Human) 642658, (Mouse) 20289

mesoderm formation tissue homeostasis endochondral ossification chondrocyte differentiation heart valve morphogenesis heart valve formation transcription, DNA-templated transcription from RNA polymerase II promoter positive regulation of cell proliferation positive regulation of gene expression cell differentiation collagen fibril organization BMP signaling pathway positive regulation of collagen biosynthetic process skeletal muscle cell differentiation tendon development tendon cell differentiation tendon formation deltoid tuberosity development negative regulation of apoptotic process negative regulation of transcription, DNA-templated positive regulation of transcription, DNA-templated positive regulation of transcription from RNA polymerase II promoter embryonic skeletal system development Sertoli cell differentiation face morphogenesis regulation of cartilage development positive regulation of cartilage development sclerotome development cellular response to mechanical stimulus cellular response to transforming growth factor beta stimulus positive regulation of gastrulation regulation of transcription, DNA-templated multicellular organism development cartilage development

Process(es)

RNA polymerase II distal enhancer sequence-specific DNA binding transcriptional activator activity, RNA polymerase II distal enhancer sequence-specific binding DNA binding bHLH transcription factor binding sequence-specific DNA binding protein heterodimerization activity E-box binding protein binding protein dimerization activity