MA1-23242

antibody from Invitrogen Antibodies

Targeting: ID3 bHLHb25, HEIR-1

Provider product page for MA1-23242

Western blot

Immunocytochemistry

Immunoprecipitation

Other assay

Antibody data

Antibody Data
Antigen structure
References [1]
Comments [0]
Validations
Immunocytochemistry [1]
Other assay [2]

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Product number: MA1-23242 - Provider product page
Provider: Invitrogen Antibodies
Product name: ID3 Monoclonal Antibody (2B11)
Antibody type: Monoclonal
Antigen: Other
Description: Recommended positive controls: Id3-transfected 293T, Id3 transfected Sol8 muscle cells and recombinant fusion protein.
Reactivity: Human, Mouse
Host: Mouse
Isotype: IgG
Antibody clone number: 2B11
Vial size: 100 µL
Concentration: 2.45 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

ID3 protein structure - MA1-23242 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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