- PA5-22924 - Invitrogen Antibodies MLXIPL antibody

Zhang P, Kumar A, Katz LS, Li L, Paulynice M, Herman MA, Scott DK
Diabetes 2015 Dec;64(12):4158-70

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ChREBP using a polyclonal antibody (Product # PA5-22924).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ChREBP using a polyclonal antibody (Product # PA5-22924).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of ChREBP using a polyclonal antibody (Product # PA5-22924).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 Depletion of ChREBPbeta attenuates glucose-stimulated beta-cell proliferation in isolated rat islet cells. A and B : Isolated rat islet cells were treated with lipid-conjugated (Accell) siRNAs and cultured in 5.5 or 15 mmol/L glucose for 96 h. Total RNA was collected and subjected to RT-PCR using primers specific for ChREBPbeta or ChREBPalpha mRNAs. C : Protein extracts were subjected to immunoblotting using an antibody against ChREBPalpha and beta-actin. D : To determine the effects of the siRNAs on ChREBP translocation, cells were treated with control or Accell siRNA against ChREBPbeta cultured in 5.5 or 15 mmol/L glucose for 32 h and were fixed and stained for insulin, ChREBP, and DNA (DAPI). E : Isolated rat islet cells were treated with Accell siRNAs and cultured in 5.5 or 15 mmol/L glucose for 96 h and fixed and stained with Ki67 and insulin. F : Quantification of the results in E , wherein at least 1,000 insulin-positive cells were counted. Results are from four different rat islet isolations. siChREBPbeta-01 and -02, siRNAs directed against ChREBPbeta; siCon, scramble control siRNA. Error bars are the SEM. * P < 0.05.
Conjugate: Horseradish Peroxidase

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 ChIP assays reveal tissue-specific recruitment of ChREBP to exon 1b ChoREs. A : INS-1-derived 832/13 cells were treated with 2 or 20 mmol/L glucose for 18 h, and cells were fixed with formaldehyde and subjected to a ChIP assay using antibodies directed against ChREBP or an IgG control. The results are presented as fold enrichment over the control IgG signal. Numerous primer pairs were chosen to scan regions of the ChREBP gene upstream of exons 1a and 1b as indicated. The results are from four to seven independent experiments. Chromatin isolated from rat islets cultured for 4 days in 15 mmol/L glucose ( B ) or from frozen mouse liver from ad libitum-fed mice ( C ) was sheared and subjected to a ChIP assay using antibodies against ChREBP or IgG as the control. Results relative to the IgG control and are from three mice and four rats. D : NIH 3T3-L1 cells were differentiated into mature adipocytes, cultured for 18 h in 20 mmol/L glucose, and processed for a quantitative ChIP assay. Results of three to four independent experiments are expressed as the signal from an antibody directed against ChREBP relative to the IgG control. Error bars are the SEM. * P < 0.05; ns, not significant.
Conjugate: Horseradish Peroxidase

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 ChREBPbeta depletion attenuates glucose-stimulated beta-cell proliferation in INS-1-derived 832/13 cells. A : INS-1-derived 832/13 cells were treated with control siRNAs (siCon) or siRNAs targeted against ChREBPbeta mRNA (Sibeta-01 and -02) and 24 h later cultured for 16 h in 2 or 20 mmol/L glucose. Immunoblotting was performed using antibodies against ChREBP (ChREBPalpha is visualized) and beta-actin (note that these siRNAs decreased ChREBPbeta mRNA but had no effect on ChREBPalpha, Fig. 1 ). pos, positive control. B : After 48 h of the siRNA treatment, cells were treated for 2 h with 2 or 20 mmol/L glucose and fixed and stained with an antibody recognizing ChREBPalpha. C : After 48 h, cells were cultured in 2 or 20 mmol/L glucose for 16 h, and BrdU was added 30 min before fixation and staining for BrdU (red) and DAPI (blue). D : Quantification of the results in C , wherein at least 1,000 cells were counted. Results are from four independent experiments. Error bars are the SEM. siChREBPbeta, siRNAs directed against ChREBPbeta. * P < 0.05.
Conjugate: Horseradish Peroxidase

PA5-22924