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Phospho-RPS6-S235 Rabbit mAb (AP1471)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat

Western blot analysis of lysates from NIH/3T3 cells using Phospho-RPS6-S235 Rabbit mAb (AP1471) at 1:3000 dilution. NIH/3T3 cells were treated by 20% FBS at 37℃ for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.

Western blot analysis of lysates from HeLa cells using Phospho-RPS6-S235 Rabbit mAb (AP1471) at 1:3000 dilution. HeLa cells were treated by 10% FBS at 37℃ for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.

Western blot analysis of lysates from PC-12 cells using Phospho-RPS6-S235 Rabbit mAb (AP1471) at 1:3000 dilution. PC-12 cells were treated by Anisomycin (100 ng/ml) at 37℃ for 24 hours.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 30s.

ABclonal:Immunohistochemistry - Phospho-RPS6-S235 Rabbit mAb (AP1471)

Immunohistochemistry analysis of paraffin-embedded Human colon carcinoma tissue using Phospho-RPS6-S235 Rabbit mAb (AP1471) at a dilution of 1:200 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse brain tissue using Phospho-RPS6-S235 Rabbit mAb (AP1471) at a dilution of 1:200 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat spleen tissue using Phospho-RPS6-S235 Rabbit mAb (AP1471) at a dilution of 1:200 (40x lens). High pressure antigen retrieval was performed with 0.01 M citrate buffer (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse liver tissue, Untreated (left) and lambda phosphatase-treated (right), using Phospho-RPS6-S235 Rabbit mAb (AP1471) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human tonsil tissue, Untreated (left) and lambda phosphatase-treated (right), using Phospho-RPS6-S235 Rabbit mAb (AP1471) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat intestine tissue, Untreated (left) and lambda phosphatase-treated (right), using Phospho-RPS6-S235 Rabbit mAb (AP1471) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

ABclonal:Immunofluorescence - Phospho-RPS6-S235 Rabbit mAb (AP1471)

Confocal imaging of HeLa cells (treated with FBS) and HeLa cells (untreated) cells using Phospho-RPS6-S235 Rabbit mAb (AP1471, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of NIH/3T3 cells (treated with FBS) and NIH/3T3 cells (untreated) cells using Phospho-RPS6-S235 Rabbit mAb (AP1471, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.

Overview

Product name	Phospho-RPS6-S235 Rabbit mAb
Catalog No.	AP1471
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC66157

Background

Ribosomes, the organelles that catalyze protein synthesis, consist of a small 40S subunit and a large 60S subunit. Together these subunits are composed of 4 RNA species and approximately 80 structurally distinct proteins. This gene encodes a cytoplasmic ribosomal protein that is a component of the 40S subunit. The protein belongs to the S6E family of ribosomal proteins. It is the major substrate of protein kinases in the ribosome, with subsets of five C-terminal serine residues phosphorylated by different protein kinases. Phosphorylation is induced by a wide range of stimuli, including growth factors, tumor-promoting agents, and mitogens. Dephosphorylation occurs at growth arrest. The protein may contribute to the control of cell growth and proliferation through the selective translation of particular classes of mRNA. As is typical for genes encoding ribosomal proteins, there are multiple processed pseudogenes of this gene dispersed through the genome.

Immunogen information

Immunogen	A synthetic phosphorylated peptide around S235 of human RPS6 (NP_001001.2).
Sequence	RLSSL
Gene ID
Swiss Prot
Synonyms	S6; eS6
Calculated MW	29kDa
Observed MW	35kDa

Applications

Reactivity	Human, Mouse, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:1000 - 1:5000 IHC-P 1:50 - 1:200 IF/ICC 1:50 - 1:200
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence
Positive samples	HeLa treated by FBS, NIH/3T3 treated by FBS, PC-12 treated by Anisomycin
Cellular location	Cytoplasm, Cytoplasmic ribonucleoprotein granule, Cytosol, Cytosolic ribosome, Endoplasmic reticulum, Nucleolus, Nucleoplasm, Nucleus, Perinuclear region of cytoplasm