44-923G

antibody from Invitrogen Antibodies

Targeting: RPS6 S6

Antibody data

Submit

Validation data

Product number: 44-923G - Provider product page
Provider: Invitrogen Antibodies
Product name: Phospho-S6 (Ser244, Ser247) Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Reactivity: Human
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RPS6 protein structure - 44-923G shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Peptide Competition. Lysates prepared from HeLa cells left untreated (1) or treated with TNF-alpha (2-5) were resolved by SDS-PAGE on a 14% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer for one hour at room temperature, and incubated with ribosomal protein S6 (pSpS244/247) antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the immunogen (3), a generic phosphoserine-containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F (ab’)2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and bands were detected using the Pierce SuperSignal™ method. The data show that only the peptide corresponding to ribosomal protein S6 (pSpS244/247) blocks the signal, verifying the specificity of the antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Peptide Competition. Lysates prepared from HeLa cells left untreated (1) or treated with TNF-alpha (2-5) were resolved by SDS-PAGE on a 14% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer for one hour at room temperature, and incubated with ribosomal protein S6 (pSpS244/247) antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the immunogen (3), a generic phosphoserine-containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F (ab)2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and bands were detected using the Pierce SuperSignal method. The data show that only the peptide corresponding to ribosomal protein S6 (pSpS244/247) blocks the signal, verifying the specificity of the antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Peptide Competition. Lysates prepared from HeLa cells left untreated (1) or treated with TNF-alpha (2-5) were resolved by SDS-PAGE on a 14% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer for one hour at room temperature, and incubated with ribosomal protein S6 (pSpS244/247) antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the immunogen (3), a generic phosphoserine-containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F (ab’)2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and bands were detected using the Pierce SuperSignal™ method. The data show that only the peptide corresponding to ribosomal protein S6 (pSpS244/247) blocks the signal, verifying the specificity of the antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1. Identification of lateral septum neurons activated by acute restraint stress. ( A ) Activity mapping of the whole brain using c-fos staining in control (naive) and restraint-stressed (RS) mice. Areas with the highest c-fos expression (green) difference are shown: the basolateral amygdala (BLA), bed nucleus of the stria terminals (BNST), insular cortex (IC), and lateral septum (LS). Insets show a 2x digital zoom of c-fos expression. Scale bars, 50 mum. ( B ) Activity-based transcriptomics (PhosphoTrap) of the lateral septum of control (naive) and restraint-stressed mice. Middle panel, plot depicting the average IP/INP value (log2) of all genes analyzed in control and restraint stress (RS) samples. Enriched genes (>2-fold; red) and depleted genes (

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Social experience with a female changes sensory activity and behavior in male mice. ( a ) Percentage of male subjects displaying pup-directed aggression, depending on the duration of cohabitation with a female (dpc). Groups are virgin mice (without contact with a female), dpc2 (2 days of cohabitation), dpc7, dpc15, and dpc22 (n = 6-12 animals). For each group, the number of aggressive individuals out of all mice assayed is represented above the bar. ( b ) Number of vomeronasal neurons immunostained for the activation marker pS6 (normalized to 50,000 mum 2 of sensory epithelium) in subjects exposed to pup odors in the same groups shown in ( a ) (n = 3-6 animals; mean + S.E.M. ; gray dots represent values for individual mice). *P < 0.05; ***P < 0.001; one-way ANOVA followed by Honestly Significantly Different post-hoc analysis; difference relative to 'virgin' group. See also Supplementary Table S1 for metrics of statistical tests and means calculated across all sections. ( c ) Representative images of VNO coronal sections subjected to pS6 immunostaining (green fluorescence) in groups shown in ( a ). DAPI nuclear staining is shown in blue. Positive control is VNO from animals exposed to predator odor (cat). ep vomeronasal sensory epithelium, lu vomeronasal organ lumen. Bar 100 mum.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 1. Differential enrichment of genes after leptin treatment. PhosphoTRAP was performed on the samples indicated below, and the RNA abundance determined by the number of RNA-seq reads was plotted in pair-wise comparisons showing ( A ) differential enrichment of genes in pS6 immunoprecipitates determined by RNA-seq in 14 d leptin treatment of ob/ob mice vs. PBS in ( A ) hypothalamus and ( B ) brainstem. TaqMan assays were then performed for the pS6-precipitated polysomes after 14 d PBS treatment of ob/ob mice after leptin treatment (for 2, 4, 7, and 14 d) of ob mice and after 14 d PBS treatment of WT mice. The genes shown are those whose level of enrichment had changed in the comparison between 14 d of leptin vs. PBS (specific genes are shown in A and B ): ( C ) Ppp1r17, ( D ) POMC, ( E ) AGRP, ( F ) Tph2, ( G ) Npw, and ( H ) CRH. Data are expressed as the ratio of fold enrichment (IP per input) for each group of mice divided by the fold enrichment (IP per input) for ob/ob mice treated with PBS for 14 d. Homogenates of pooled hypothalami from 15 to 20 mice are used for each group.