PA5-37733

antibody from Invitrogen Antibodies

Targeting: RPS6KB1 p70(S6K)-alpha, PS6K, S6K, S6K1, STK14A

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-37733

Provider

Invitrogen Antibodies

Product name

Phospho-p70 S6 Kinase (Thr421) Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

A suggested positive control for Western blot is Jurkat cells; suggested positive control for IHC is human breast carcinoma; suggested positive control for ICC/IF is MCF-7 cells.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

1 mg/mL

Storage

-20°C

References

Submitted references

Increased mTOR activity and metabolic efficiency in mouse and human cells containing the African-centric tumor-predisposing p53 variant Pro47Ser.

Gnanapradeepan K, Leu JI, Basu S, Barnoud T, Good M, Lee JV, Quinn WJ, Kung CP, Ahima R, Baur JA, Wellen KE, Liu Q, Schug ZT, George DL, Murphy ME
eLife 2020 Nov 10;9

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of extracts from Jurkat cells, untreated (Lane 1) or treated with GM-CSF (Lane 2), using p70 S6 Kinase (pThr421) polyclonal antibody (Product # PA5-37733).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunocytochemical analysis of Phospho-p70 S6 Kinase (Thr421) in methanol-fixed MCF cells, using Phospho-p70 S6 Kinase (Thr421) Polyclonal Antibody (Product # PA5-37733).

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of Phospho-p70 S6 Kinase (Thr421) in paraffin-embedded Human breast carcinoma tissue using Phospho-p70 S6 Kinase (Thr421) Polyclonal Antibody (Product # PA5-37733) (left) or the same antibody preincubated with blocking peptide (right).

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 1. Increased markers of mTOR activity in S47 cells and tissues. ( A ) Western blot analyses reveal higher phospho-S6K1 expression in S47 LCLs and S47 MEFs; the latter were obtained from two separate embryos per genotype. ( B ) Densitometry quantification of phospho-S6K1 protein expression in WT and S47 MEFs from four independent experiments; all values normalized to total S6K1. Error bars represent standard error, (*) p value < 0.05. ( C ) Whole cell lysates were extracted from three WT and three S47 mouse lungs and analyzed by western blot for the proteins indicated. Pairs 1 and 3 are lungs isolated from male mice, pair 2 is lungs isolated from female mice. Densitometry quantification of phospho-S6K1 and phospho-mTOR was performed and normalized to total S6K1 and total mTOR protein expression, respectively. ( D ) Whole cell lysates were extracted from WT and S47 mouse skeletal muscle and analyzed as described above. Densitometry quantification of phospho-S6, phospho-S6K1, phospho-mTOR was performed and normalized to total S6, total S6K1 and total mTOR protein expression, respectively. ( E ) Immunohistochemical analysis of hematoxylin and eosin (H and E), phospho-mTOR and phospho-S6K1 in WT and S47 mouse lung and skeletal tissue. Data are representative of n = 4 mice per genotype. Scale bar represents 100 mum. ( F ) WT and S47 MEFs were grown in glucose-free media for 16 hr, then in media containing 4.5 g/L glucose. Samples were collected at indicated time points and a