Product: Phospho-CaMK4 (Thr200) Antibody
Catalog: AF3460
Description: Rabbit polyclonal antibody to Phospho-CaMK4 (Thr200)
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Horse, Sheep, Dog, Chicken, Xenopus
Mol.Wt.: 60kDa; 52kD(Calculated).
Uniprot: Q16566
RRID: AB_2834898

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 100ul $280 In stock
 200ul $350 In stock

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Zebrafish(100%), Horse(100%), Sheep(100%), Dog(100%), Chicken(100%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
Phospho-CaMK4 (Thr200) Antibody detects endogenous levels of CaMK4 only when phosphorylated at Threonine 200.
RRID:
AB_2834898
Cite Format: Affinity Biosciences Cat# AF3460, RRID:AB_2834898.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

Brain Ca(2+) calmodulin dependent protein kinase type 4; Brain Ca(2+) calmodulin dependent protein kinase type IV; Brain Ca++-calmodulin dependent protein kinase type IV; Calcium / calmodulin dependent protein kinase type 4 catalytic chain; Calcium / calmodulin dependent protein kinase type IV catalytic chain; Calcium/calmodulin dependent protein kinase IV; Calcium/calmodulin dependent protein kinase type IV; Calcium/calmodulin-dependent protein kinase type IV; CAM kinase 4; CAM kinase GR; CAM kinase IV; CAM kinase-GR; CaMK 4; CAMK GR; CaMK IV; Camk4; CaMKGR; IV; KCC4_HUMAN; MGC36771;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
Q16566 KCC4_HUMAN:

Expressed in brain, thymus, CD4 T-cells, testis and epithelial ovarian cancer tissue.

Description:
The product of this gene belongs to the serine/threonine protein kinase family, and to the Ca(2+)/calmodulin-dependent protein kinase subfamily. This enzyme is a multifunctional serine/threonine protein kinase with limited tissue distribution, that has been implicated in transcriptional regulation in lymphocytes, neurons and male germ cells.
Sequence:
MLKVTVPSCSASSCSSVTASAAPGTASLVPDYWIDGSNRDALSDFFEVESELGRGATSIVYRCKQKGTQKPYALKVLKKTVDKKIVRTEIGVLLRLSHPNIIKLKEIFETPTEISLVLELVTGGELFDRIVEKGYYSERDAADAVKQILEAVAYLHENGIVHRDLKPENLLYATPAPDAPLKIADFGLSKIVEHQVLMKTVCGTPGYCAPEILRGCAYGPEVDMWSVGIITYILLCGFEPFYDERGDQFMFRRILNCEYYFISPWWDEVSLNAKDLVRKLIVLDPKKRLTTFQALQHPWVTGKAANFVHMDTAQKKLQEFNARRKLKAAVKAVVASSRLGSASSSHGSIQESHKASRDPSPIQDGNEDMKAIPEGEKIQGDGAQAAVKGAQAELMKVQALEKVKGADINAEEAPKMVPKAVEDGIKVADLELEEGLAEEKLKTVEEAAAPREGQGSSAVGFEVPQQDVILPEY

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Sheep
100
Dog
100
Xenopus
100
Zebrafish
100
Chicken
100
Bovine
0
Rabbit
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q16566 As Substrate

Site PTM Type Enzyme
S12 Phosphorylation Q8N5S9 (CAMKK1) , Q16566 (CAMK4)
S13 Phosphorylation Q8N5S9 (CAMKK1) , Q16566 (CAMK4)
Y32 Phosphorylation
S50 Phosphorylation
T57 O-Glycosylation
S58 O-Glycosylation
K66 Ubiquitination
K70 Ubiquitination
K75 Ubiquitination
K133 Ubiquitination
S137 O-Glycosylation
K166 Ubiquitination
K182 Ubiquitination
S189 O-Glycosylation
S189 Phosphorylation
K190 Acetylation
K190 Ubiquitination
K199 Ubiquitination
T200 Phosphorylation Q8N5S9 (CAMKK1) , Q96RR4 (CAMKK2)
K279 Ubiquitination
K303 Ubiquitination
K316 Ubiquitination
K331 Ubiquitination
S341 Phosphorylation Q16566 (CAMK4)
S343 Phosphorylation
S344 O-Glycosylation
S344 Phosphorylation
S345 O-Glycosylation
S345 Phosphorylation
S348 Phosphorylation
S352 Phosphorylation
K354 Ubiquitination
S356 O-Glycosylation
S356 Phosphorylation
S360 Phosphorylation Q16566 (CAMK4)
K404 Ubiquitination
K440 Ubiquitination
K442 Ubiquitination

PTMs - Q16566 As Enzyme

Substrate Site Source
P14866 (HNRNPL) S544 Uniprot
P16220 (CREB1) S133 Uniprot
P16949 (STMN1) S16 Uniprot
P29475-1 (NOS1) S852 Uniprot
P53667 (LIMK1) T508 Uniprot
P56524-1 (HDAC4) S467 Uniprot
P56524-1 (HDAC4) S632 Uniprot
Q04206 (RELA) S536 Uniprot
Q16566 (CAMK4) S12 Uniprot
Q16566 (CAMK4) S13 Uniprot
Q16566 (CAMK4) S341 Uniprot
Q16566 (CAMK4) S360 Uniprot
Q92793-1 (CREBBP) S302 Uniprot
Q99623 (PHB2) S91 Uniprot
Q9UQL6 (HDAC5) S259 Uniprot
Q9UQL6 (HDAC5) S498 Uniprot
Q9UQM7-2 (CAMK2A) S332 Uniprot

Research Backgrounds

Function:

Calcium/calmodulin-dependent protein kinase that operates in the calcium-triggered CaMKK-CaMK4 signaling cascade and regulates, mainly by phosphorylation, the activity of several transcription activators, such as CREB1, MEF2D, JUN and RORA, which play pivotal roles in immune response, inflammation, and memory consolidation. In the thymus, regulates the CD4(+)/CD8(+) double positive thymocytes selection threshold during T-cell ontogeny. In CD4 memory T-cells, is required to link T-cell antigen receptor (TCR) signaling to the production of IL2, IFNG and IL4 (through the regulation of CREB and MEF2). Regulates the differentiation and survival phases of osteoclasts and dendritic cells (DCs). Mediates DCs survival by linking TLR4 and the regulation of temporal expression of BCL2. Phosphorylates the transcription activator CREB1 on 'Ser-133' in hippocampal neuron nuclei and contribute to memory consolidation and long term potentiation (LTP) in the hippocampus. Can activate the MAP kinases MAPK1/ERK2, MAPK8/JNK1 and MAPK14/p38 and stimulate transcription through the phosphorylation of ELK1 and ATF2. Can also phosphorylate in vitro CREBBP, PRM2, MEF2A and STMN1/OP18.

PTMs:

Phosphorylated by CaMKK1 and CaMKK2 on Thr-200. Dephosphorylated by protein phosphatase 2A. Autophosphorylated on Ser-12 and Ser-13.

Glycosylation at Ser-189 modulates the phosphorylation of CaMK4 at Thr-200 and negatively regulates its activity toward CREB1 in basal conditions and during early inomycin stimulation.

Subcellular Location:

Cytoplasm. Nucleus.
Note: Localized in hippocampal neuron nuclei. In spermatids, associated with chromatin and nuclear matrix (By similarity).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed in brain, thymus, CD4 T-cells, testis and epithelial ovarian cancer tissue.

Subunit Structure:

Monomer (By similarity). Interacts with protein phosphatase 2A (PPP2CA/PPP2CB); the interaction is mutually exclusive with binding to Ca(2+)/calmodulin.

Family&Domains:

The autoinhibitory domain overlaps with the calmodulin binding region and interacts in the inactive folded state with the catalytic domain as a pseudosubstrate.

Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CaMK subfamily.

Research Fields

· Environmental Information Processing > Signal transduction > Calcium signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > cAMP signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Apelin signaling pathway.   (View pathway)

· Human Diseases > Substance dependence > Amphetamine addiction.

· Human Diseases > Substance dependence > Alcoholism.

· Organismal Systems > Aging > Longevity regulating pathway.   (View pathway)

· Organismal Systems > Development > Osteoclast differentiation.   (View pathway)

· Organismal Systems > Nervous system > Long-term potentiation.

· Organismal Systems > Nervous system > Neurotrophin signaling pathway.   (View pathway)

· Organismal Systems > Nervous system > Cholinergic synapse.

· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

· Organismal Systems > Endocrine system > Aldosterone synthesis and secretion.

References

1). Calcium Channel Blocker Nifedipine Suppresses Colorectal Cancer Progression and Immune Escape by Preventing NFAT2 Nuclear Translocation.

Application: WB    Species: Human    Sample: SW620 cells

Figure 3. The Calcium-Dependent Gene NFAT2 Is Activated through Dephosphorylation and Transportation to the Nucleus (A) IF assay was used to visualize the alteration of NFAT2 in SW620 cells treated with NIFE (10 mM) or the NFAT2 vector. (B) IF assay was used to visualize the alteration in NFAT2 in SW620 cells treated with NFAT2 short hairpin RNA (shRNA) or BAY (10 mM). (C) IF assays were performed with subcutaneous tumor slices from mice treated with NIFE or BAY to visualize the subcellular localization of NFAT2. (D) After treating SW620 cells with NIFE or BAY, IF assays were used to visualize the subcellular localization of NFAT2 and phosphorylated NFAT2 (p-NFAT2). (E) IF assay was used to visualize the Ca2+ level and NFAT2 or p-NFAT2 level in normal tissues and tumors. Images in the same line were from the same magnification. Blue light represents the basic light of Ca2+, whereas green light represents the elevated Ca2+ levels. Red light represents the level of NFAT2. (F) Upper panel: detection of total protein changes in SW620 cells after treatment with NIFE or transfection with siRNA-CAMK-II by WB. Lower panel: proteins contained in cytoplasmic and nuclear compartments of SW620 cells after treatment with NIFE or BAY were separated for WB analysis. (G) After treating SW620 cells with CsA (10 mM), VIVIT (10 mM), and FK506 (10 ng/mL) and subsequently separating the nuclear and cytoplasmic proteins, WB analysis was used to detect the level of NFAT2 in the nucleus and p-NFAT2 in the cytoplasm. (H) Representative images of IHC staining analysis of NFAT2 expression in CRC tissues and adjacent nontumor tissues. The bar chart on the right represents the percentage of high and low NFAT2 expression cases in normal and CRC tissues. (I) IHC analysis of NFAT2 and p-NFAT2 expression in nonmetastatic and metastatic CRC tissues. The bar chart on the right represents the percentage of high and low NFAT2 or p-NFAT2 expression cases in nonmetastatic CRC and metastatic CRC tissues.

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