phospho-NFATc2 (Ser330) Polyclonal Antibody

$Molt4 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with phospho-NFATc2 (Ser330) Polyclonal Antibody(bs-5516R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

Molt4 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with phospho-NFATc2 (Ser330) Polyclonal Antibody(bs-5516R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

$Lane 1: Mouse Thymus tissue lysates; Lane 2: Mouse Spleen tissue lysates; Lane 3: Human Raji cell lysates; Lane 4: Human Jurkat cell lysates; Lane 5: Human U251 cell lysates probed with phospho-NFATc2 (Ser330) Polyclonal Antibody, Unconjugated (bs-5516R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.$

Lane 1: Mouse Thymus tissue lysates; Lane 2: Mouse Spleen tissue lysates; Lane 3: Human Raji cell lysates; Lane 4: Human Jurkat cell lysates; Lane 5: Human U251 cell lysates probed with phospho-NFATc2 (Ser330) Polyclonal Antibody, Unconjugated (bs-5516R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

$HUVEC cells were\u00a0fixed with 4% PFA for 10min at room temperature,permeabilized with\u00a090% ice-cold methanol for 20 min at -20\u2103,\u00a0and incubated in 5% BSA blocking buffer\u00a0for 30 min at room temperature. Cells were then stained with phospho-NFATc2 (Ser330) Polyclonal Antibody(bs-5516R)at 1:100 dilution\u00a0in blocking buffer and\u00a0incubated for 30 min at\u00a0room temperature,\u00a0washed twice with 2%BSA in PBS,\u00a0followed by\u00a0secondary antibody incubation\u00a0for 40 min\u00a0at\u00a0room temperature. Acquisitions of 20,000 events were performed.\u00a0Cells stained with primary antibody\u00a0(green), and isotype control (orange).$

HUVEC cells were\u00a0fixed with 4% PFA for 10min at room temperature,permeabilized with\u00a090% ice-cold methanol for 20 min at -20\u2103,\u00a0and incubated in 5% BSA blocking buffer\u00a0for 30 min at room temperature. Cells were then stained with phospho-NFATc2 (Ser330) Polyclonal Antibody(bs-5516R)at 1:100 dilution\u00a0in blocking buffer and\u00a0incubated for 30 min at\u00a0room temperature,\u00a0washed twice with 2%BSA in PBS,\u00a0followed by\u00a0secondary antibody incubation\u00a0for 40 min\u00a0at\u00a0room temperature. Acquisitions of 20,000 events were performed.\u00a0Cells stained with primary antibody\u00a0(green), and isotype control (orange).

$HUVEC cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (phospho-NFATc2 (Ser330)) polyclonal Antibody, Unconjugated (bs-5516R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.$

HUVEC cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (phospho-NFATc2 (Ser330)) polyclonal Antibody, Unconjugated (bs-5516R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Applications

WB
IHC-P
IHC-F
IF
Flow-Cyt

Reactivity

Human
Mouse
Rat

Predicted Reactivity

Dog
Cow
Pig
Horse

Overview
Catalog #	bs-5516R
Product Name	phospho-NFATc2 (Ser330) Polyclonal Antibody
Applications	WB, IHC-P, IHC-F, IF, Flow-Cyt
Reactivity	Human, Mouse, Rat
Predicted Reactivity	Dog, Cow, Pig, Horse
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic phosphopeptide derived from human NFATc2 around the phosphorylation site of Ser330
Modification Site	Ser330
Clonality	Polyclonal
Clone #	#REF!
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4_. Store at -20_ for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	4773
Subcellular location	Cytoplasm, Nucleus
Synonyms	NFATC2phospho S330; AI607462; KIAA0611; NF ATp; NF-ATc2; NFAT 1; NFAT pre existing subunit; NFAT transcription complex, preexisting component; NFAT1; NFAT1-D; NFATc2; NFATp; Nuclear factor of activated T cells cytoplasmic 2; Nuclear factor of activated T cells cytoplasmic calcineurin dependent 2; Nuclear factor of activated T cells pre-existing component; T cell transcription factor NFAT.
Background	Nuclear factor of activated T cells (NFAT) is a family of transcription factors implicated in multiple biological processes including cytokine gene expression, cardiac hypertrophy and adipocyte differentiation. NFAT1 (also known as NFATc2 or NFATp) is a member of this family that is regulated by the calcium-dependent phosphatase calcineurin. When calcineurin is activated by calcium it dephosphorylates multiple residues in the regulatory domain of NFAT1, leading to its translocation to the nucleus and activation of its transcriptional activity. Once in the nucleus, NFAT proteins act synergistically with the AP-1 transcription factor complex to regulate the expression of multiple genes. Serine 54 in mouse NFAT1 has been shown to be important in the regulation of its transcriptional activity.
Application Dilution
WB	1:300-5000
IHC-P	ELISA1:500-1000
IHC-F	IHC-P1:200-400
IF
Flow-Cyt