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Phospho-ACLY-S455 Rabbit pAb (AP0779)

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 C6 cells using Phospho-ACLY-S455 Rabbit pAb (AP0779) at 1:400 dilution. C6 cells were treated by Insulin (100 ng/mL) 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: 60s.

Western blot analysis of lysates from NIH/3T3 cells using Phospho-ACLY-S455 Rabbit pAb (AP0779) at 1:400 dilution. NIH/3T3 cells were treated by Insulin (200 nM) 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: 60s.

Western blot analysis of lysates from HeLa cells using Phospho-ACLY-S455 Rabbit pAb (AP0779) at 1:400 dilution. HeLa cells were treated by Insulin (50 nM) 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: 60s.

ABclonal:Immunohistochemistry - Phospho-ACLY-S455 Rabbit pAb (AP0779)

Immunohistochemistry analysis of paraffin-embedded Rat pancreas using Phospho-ACLY-S455 Rabbit pAb (AP0779) at dilution of 1:100 (40x lens). Microwave antigen retrieval performed with 0.01M Tris/EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse brain using Phospho-ACLY-S455 Rabbit pAb (AP0779) at dilution of 1:100 (40x lens). Microwave antigen retrieval performed with 0.01M Tris/EDTA Buffer (pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse pancreas using Phospho-ACLY-S455 Rabbit pAb (AP0779) at dilution of 1:100 (40x lens). Microwave antigen retrieval performed with 0.01M Tris/EDTA Buffer (pH 9.0) prior to IHC staining.

ABclonal:Immunoprecipitation - Phospho-ACLY-S455 Rabbit pAb (AP0779)

Immunoprecipitation analysis of 200 μg extracts of NIH/3T3 cells, using 3 μg Phospho-ACLY-S455 pAb (AP0779). Western blot was performed from the immunoprecipitate using Phospho-ACLY-S455 pAb (AP0779) at a dilution of 1:1000. NIH/3T3 cells were treated by Insulin (100 nM) at 37℃ for 10 minutes after serum-starvation overnight.

Overview

Product name	Phospho-ACLY-S455 Rabbit pAb
Catalog No.	AP0779
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG

Background

ATP citrate lyase is the primary enzyme responsible for the synthesis of cytosolic acetyl-CoA in many tissues. The enzyme is a tetramer (relative molecular weight approximately 440, 000) of apparently identical subunits. It catalyzes the formation of acetyl-CoA and oxaloacetate from citrate and CoA with a concomitant hydrolysis of ATP to ADP and phosphate. The product, acetyl-CoA, serves several important biosynthetic pathways, including lipogenesis and cholesterogenesis. In nervous tissue, ATP citrate-lyase may be involved in the biosynthesis of acetylcholine. Multiple transcript variants encoding distinct isoforms have been identified for this gene.

Immunogen information

Immunogen	A synthetic phosphorylated peptide around S455 of human ACLY (NP_001087.2).
Sequence	RTASFS
Gene ID
Swiss Prot
Synonyms	ACL; ATPCL; CLATP; Phospho-ACLY-S455
Calculated MW	121kDa
Observed MW	125kDa

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 1:100 - 1:500 IHC-P 1:50 - 1:100 IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunoprecipitation
Positive samples	C6 treated by Insulin, NIH/3T3 treated by Insulin, HeLa treated by Insulin
Cellular location	Cytoplasm.