PA5-40553

antibody from Invitrogen Antibodies

Targeting: HEY1 bHLHb31, CHF-2, CHF2, HERP2, HESR-1, HESR1, HRT-1

Western blot

Immunohistochemistry

Antibody data

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Validation data

Product number: PA5-40553 - Provider product page
Provider: Invitrogen Antibodies
Product name: HEY1 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: Peptide sequence: PQNGHGNAGT TASPTEPHHQ GRLGSAHPEA PALRAPPSGS FGPVLPVVTS Sequence homology: Cow: 79%; Dog: 86%; Guinea Pig: 93%; Horse: 93%; Human: 100%; Mouse: 93%; Rabbit: 79%; Rat: 93%
Reactivity: Human
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 0.5 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

HEY1 protein structure - PA5-40553 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 6 Effect of flutamide (F) and EDS on Hey1 , Hes1 , and Hes5 expression in rat testis. (A - C) Relative expression of Hey1, Hes1, and Hes5 mRNAs was determined using real-time RT-PCR analysis. The histograms are the quantitative representation of data of three independent analyses, each in triplicate ( n = 6 each group). The expression values of the individual genes were normalized to the mean expression of the reference genes ( Rn18s , B2m and Actb ) as an internal control. Relative quantification (RQ) is expressed as mean +- SD. Significant differences from control values are denoted as ** p < 0.01 and *** p < 0.001. (A'- C') Relative protein expression of HEY1, HES1 and HES5 was determined using western blot. The histograms are the quantitative representation after densitometry of data (mean +- SD) of three independent analyses ( n = 6 each group). The relative level of studied protein was normalized against its corresponding actin data point. The protein levels within the control group were arbitrarily set at 1. Significant differences from control values are denoted as * p < 0.05, ** p < 0.01, and *** p < 0.001

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 The effect of testosterone on Notch1 , Hey1 and Hes1 mRNA, and Notch1 intracellular domain (N1ICD), HEY1 and HES1 protein expression in TM4 and 15P-1 Sertoli cell lines. Cells were treated with a vehicle (control, C), 10 -8 M, 10 -7 M or 10 -6 M testosterone (T) for 24 h ( a , b ) Relative expression of mRNAs (RQ) was determined using quantitative real-time RT-PCR analysis. The expression values of the individual genes were normalized to the mean expression of Rn18s , B2m , Gapdh and Actb . ( c , d ) Western blot detection of the proteins. The relative level of studied proteins was normalized to beta-actin. The protein levels within the control group were arbitrarily set at 1. The histograms are the quantitative representation of data (mean +- SD) of three independent experiments, each in triplicate. Significant differences from control values are denoted as * p < 0.05, ** p < 0.01, and *** p < 0.001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 The effect of AR and ZIP9 knockdown on Hey1 mRNA and HEY1 protein expression in TM4 ( a , c ) and 15P-1 ( b , d ) Sertoli cells. Cells were treated with transfection reagent alone (C), transfection reagent + 5 x 10 -8 M non-targeting siRNA (negative control, NT), transfection reagent + 5 x 10 -8 M AR siRNA (AR-Kd) or ZIP9 siRNA (ZIP9-Kd). After 24 h 10 -8 M T or vehicle was added to the culture. ( a , b ) Relative expression of mRNAs (RQ) was determined using real-time RT-PCR analysis. The expression values of the individual genes were normalized to the mean expression of Rn18s , B2m , Gapdh and Actb . ( c , d ) Western blot detection of the proteins. The relative level of studied protein was normalized to beta-actin. The protein levels within the control group were arbitrarily set at 1. The histograms are the quantitative representation of data (mean +- SD) of three independent experiments, each in triplicate. Significant differences from control values are denoted as * p < 0.05, ** p < 0.01, and *** p < 0.001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 Immunofluorescence analysis of HEY1 expression in TM4 ( a , c ) and 15P-1 ( b , d ) cells following AR or ZIP9 knockdown. Cells were transfected with transfection reagent alone (C), transfection reagent + 5 x 10 -8 M AR siRNA (AR-Kd) or ZIP9 siRNA (ZIP9-Kd). After 24 h 10 -8 M T or vehicle was added to the culture. Cells were fixed after a 24 h incubation. ( a , b ) Representative microphotographs of TM4 and 15P-1 cultures. Arrows indicate positive signal. Sertoli cell nuclei were stained with DAPI (blue). Right panels in controls (C) represent the merged images. Scale bar = 25 um. ( c , d ) The fluorescence intensity was quantified using ImageJ and displayed in corrected total cell fluorescence (CTCF). Histograms represent the mean +- SD. Significant differences from control values are denoted as ** p < 0.01, and *** p < 0.001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 The effect of hydroxyflutamide (HF) and bicalutamide (Bic) on Hey1 mRNA and HEY1 protein expression in TM4 ( a , d , g , j ), 15P-1 ( b , e , h , k ), and primary (PSC; c , f , j , l ) Sertoli cells. Cells were treated with 10 -8 M testosterone (T), 10 -4 HF, HF + T, 10 -6 M Bic, Bic + T or vehicle (C) for 24 h. ( a - c ) Relative expression of mRNAs (RQ) was determined using real-time RT-PCR analysis. The expression values of the individual genes were normalized to the mean expression of Rn18s , B2m , Gapdh and Actb . ( d - f ) Western blot detection of the proteins. The relative level of studied protein was normalized to beta-actin. The protein levels within the control group were arbitrarily set at 1. The histograms are the quantitative representation of data (mean +- SD) of three independent experiments, each in triplicate. ( g - i ) Immunofluorescence analysis of HEY1 expression. Arrows indicate positive signal. Scale bar = 10 um. ( j - l ) The fluorescence intensity was quantified using ImageJ and displayed in corrected total cell fluorescence (CTCF). Histograms represent the mean +- SD. Significant differences from control values are denoted as * p < 0.05, ** p < 0.01, and *** p < 0.001.