Phospho-CaMKK2 (Ser511) Antibody - #AF4487

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Product: Phospho-CaMKK2 (Ser511) Antibody
Catalog: AF4487
Description: Rabbit polyclonal antibody to Phospho-CaMKK2 (Ser511)
Application: WB IHC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken
Mol.Wt.: 68kDa; 65kD(Calculated).
Uniprot: Q96RR4
RRID: AB_2844538

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 100ul $350 In stock
 200ul $450 In stock

Lead Time: Same day delivery

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Related Downloads
MS Report
HPLC Report
MSDS
Data Sheet
COA
Protocols
WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200
*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%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%)
Clonality:
Polyclonal
Specificity:
Phospho-CaMKK2 (Ser511) Antibody detects endogenous levels of CaMKK2 only when phosphorylated at Ser511.
RRID:
AB_2844538
Cite Format: Affinity Biosciences Cat# AF4487, RRID:AB_2844538.
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

Calcium/calmodulin dependent protein kinase beta; Calcium/calmodulin dependent protein kinase kinase 2; Calcium/calmodulin dependent protein kinase kinase 2 beta; Calcium/calmodulin dependent protein kinase kinase beta; Calcium/calmodulin-dependent protein kinase kinase 2; Calcium/calmodulin-dependent protein kinase kinase beta; CaM kinase kinase beta; CaM KK beta; CaM-kinase kinase 2; CaM-kinase kinase beta; CaM-KK 2; CaM-KK beta; CaMKK 2; CAMKK; CaMKK beta; CAMKK beta protein; Camkk2; CAMKKB; KKCC2_HUMAN;

Immunogens

Immunogen:
Uniprot:

Q96RR4(KKCC2_HUMAN) >>Visit HPA database.

Gene(ID):
CAMKK2(10645)
Expression:
Q96RR4 KKCC2_HUMAN:

Ubiquitously expressed with higher levels in the brain. Intermediate levels are detected in spleen, prostate, thyroid and leukocytes. The lowest level is in lung.

Sequence:
MSSCVSSQPSSNRAAPQDELGGRGSSSSESQKPCEALRGLSSLSIHLGMESFIVVTECEPGCAVDLGLARDRPLEADGQEVPLDTSGSQARPHLSGRKLSLQERSQGGLAAGGSLDMNGRCICPSLPYSPVSSPQSSPRLPRRPTVESHHVSITGMQDCVQLNQYTLKDEIGKGSYGVVKLAYNENDNTYYAMKVLSKKKLIRQAGFPRRPPPRGTRPAPGGCIQPRGPIEQVYQEIAILKKLDHPNVVKLVEVLDDPNEDHLYMVFELVNQGPVMEVPTLKPLSEDQARFYFQDLIKGIEYLHYQKIIHRDIKPSNLLVGEDGHIKIADFGVSNEFKGSDALLSNTVGTPAFMAPESLSETRKIFSGKALDVWAMGVTLYCFVFGQCPFMDERIMCLHSKIKSQALEFPDQPDIAEDLKDLITRMLDKNPESRIVVPEIKLHPWVTRHGAEPLPSEDENCTLVEVTEEEVENSVKHIPSLATVILVKTMIRKRSFGNPFEGSRREERSLSAPGNLLTKKPTRECESLSELKEARQRRQPPGHRPAPRGGGGSALVRGSPCVESCWAPAPGSPARMHPLRPEEAMEPE

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

PTMs - Q96RR4 As Substrate

Site PTM Type Enzyme
S2 Acetylation
K32 Ubiquitination
T85 Phosphorylation Q96RR4 (CAMKK2)
S95 Phosphorylation
K98 Ubiquitination
S100 Phosphorylation
S114 Phosphorylation
S125 Phosphorylation
Y128 Phosphorylation
S129 Phosphorylation Q00535 (CDK5) , P49841 (GSK3B) , P49840 (GSK3A)
S132 Phosphorylation
S133 Phosphorylation P49840 (GSK3A) , P49841 (GSK3B) , Q00535 (CDK5)
S136 Phosphorylation
S137 Phosphorylation P49841 (GSK3B) , P49840 (GSK3A) , Q00535 (CDK5)
T145 Phosphorylation
S148 Phosphorylation
K173 Ubiquitination
Y183 Phosphorylation
Y190 Phosphorylation
K194 Ubiquitination
Y234 Phosphorylation
K241 Ubiquitination
K242 Ubiquitination
K307 Ubiquitination
K314 Ubiquitination
K327 Ubiquitination
T350 Phosphorylation
K403 Ubiquitination
K420 Ubiquitination
K429 Ubiquitination
K441 Ubiquitination
T483 Phosphorylation Q96RR4 (CAMKK2)
K488 Ubiquitination
S495 Phosphorylation
S509 Phosphorylation
S511 Phosphorylation P53355 (DAPK1)
S572 Phosphorylation

PTMs - Q96RR4 As Enzyme

Substrate Site Source
P54646 (PRKAA2) T172 Uniprot
Q13131 (PRKAA1) T183 Uniprot
Q14012 (CAMK1) T177 Uniprot
Q16566 (CAMK4) T200 Uniprot
Q8IU85 (CAMK1D) T180 Uniprot
Q96EB6 (SIRT1) S27 Uniprot
Q96EB6 (SIRT1) S47 Uniprot
Q96RR4 (CAMKK2) T85 Uniprot
Q96RR4 (CAMKK2) T483 Uniprot

Research Backgrounds

Function:

Calcium/calmodulin-dependent protein kinase belonging to a proposed calcium-triggered signaling cascade involved in a number of cellular processes. Isoform 1, isoform 2 and isoform 3 phosphorylate CAMK1 and CAMK4. Isoform 3 phosphorylates CAMK1D. Isoform 4, isoform 5 and isoform 6 lacking part of the calmodulin-binding domain are inactive. Efficiently phosphorylates 5'-AMP-activated protein kinase (AMPK) trimer, including that consisting of PRKAA1, PRKAB1 and PRKAG1. This phosphorylation is stimulated in response to Ca(2+) signals (By similarity). Seems to be involved in hippocampal activation of CREB1 (By similarity). May play a role in neurite growth. Isoform 3 may promote neurite elongation, while isoform 1 may promoter neurite branching.

PTMs:

Autophosphorylated and phosphorylated by PKA. Each isoform may show a different pattern of phosphorylation.

Subcellular Location:

Nucleus. Cytoplasm. Cell projection>Neuron projection.
Note: Predominantly nuclear in unstimulated cells, relocalizes into cytoplasm and neurites after forskolin induction.

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

Ubiquitously expressed with higher levels in the brain. Intermediate levels are detected in spleen, prostate, thyroid and leukocytes. The lowest level is in lung.

Subunit Structure:

Interacts with calmodulin.

Family&Domains:

The autoinhibitory domain overlaps with the calmodulin binding region and may be involved in intrasteric autoinhibition.

The RP domain (arginine/proline-rich) is involved in the recognition of CAMKI and CAMK4 as substrates.

Belongs to the protein kinase superfamily. Ser/Thr protein kinase family.

Research Fields

· Cellular Processes > Transport and catabolism > Autophagy - animal.   (View pathway)

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

· Human Diseases > Substance dependence > Alcoholism.

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

· Organismal Systems > Endocrine system > Adipocytokine signaling pathway.

· Organismal Systems > Endocrine system > Oxytocin signaling pathway.

References

1). NCAPD2 inhibits autophagy by regulating Ca2+/CAMKK2/AMPK/mTORC1 pathway and PARP-1/SIRT1 axis to promote colorectal cancer. CANCER LETTERS (PubMed: 34229059) [IF=9.7]

Application: WB    Species: Mice    Sample: CRC cells

Fig. 2. NCAPD2 inhibited cell autophagy and disrupted autophagic flux via Ca 2+ /CAMKK2/AMPK/mTORC1 pathway. (A) Western blot analyses for phosphorylated mTOR (p-mTOR, S2448), phosphorylated p70S6K (p-p70S6K, T389/412), phosphorylated 4E-BP1 (p-4E-BP1, T70) and phosphorylated AKT (p-AKT, S473) in CRCC cells with different treatments as indicated. (B) Western blot of indicated proteins in cells treated with mTORC1 inhibitor Rapamycin (3 nM, 24h). (C) Immuno- fluorescence staining of LC3II (red) and P62 (red) in CRC cells with different treatments as indicated. Merged images represented overlays of LC3II or P62 and nuclear staining by DAPI (blue). (D) Intracellular Ca 2+ levels were analyzed by flow cytometry after staining with the fluorescent probe Fluo-3, AM in CRC cells. (E) Representative Western blot gel documents of phosphorylated CAMKK2(S511), phosphorylated AMPK(T172), phosphorylated mTOR(S2448), Beclin, ATG5, P62, LC3II in CRC cells with different treatments. (F) Western blots of indicated proteins in cells treated with an inhibitor of microsomal Ca 2+ -ATPase Thapsigargin (1 μ M, 6h) and Ca 2+ chelator BAPTA-AM (10 μ M, 12h) respectively. Results are shown as mean ± s.d, *P < 0.05, **P < 0.01, ***P < 0.001, based on Student’s t-test. . (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
2). Caffeic acid dimethyl ether alleviates alcohol-induced hepatic steatosis via microRNA-378b-mediated CaMKK2-AMPK pathway. Bioengineered (PubMed: 35481488) [IF=4.9]

Application: WB    Species: mouse    Sample: livers

Figure 2: |Effect of CADE on miR-378b-CaMKK2 signal. (a) The expression level of miR-378b in liver tissues. (b) CaMKK2 mRNA expression. (c) Protein expression levels of CaMKK2 and p-CaMKK2. All data are expressed as means ± SD.
3). Ephedra Herb extract ameliorates adriamycin-induced nephrotic syndrome in rats via the CAMKK2/AMPK/mTOR signaling pathway. Chinese Journal of Natural Medicines [IF=4.6]
4). L-theanine prevents progression of nonalcoholic hepatic steatosis by regulating hepatocyte lipid metabolic pathways via the CaMKKβ-AMPK signaling pathway. Nutrition & Metabolism (PubMed: 35428314) [IF=4.5]

Application: WB    Species: Human    Sample: HepG2 cells

Fig. 7 L-theanine regulates hepatocyte lipid metabolic pathways via the CaMKKβ-AMPK signaling pathway. HepG2 cells were treated with 500 μM OA for 24 h with or without pretreated L-theanine for 2 h. A Western blot analysis showing protein expression of p-CaMKKβ and p-AMPK in HepG2 cells. HepG2 cells were pretreated with 10 μM STO-609 for 2 h, then added L-theanine co-incubated for 2 h, finally, OA were added co-incubated for 24 h. B Abrogating effect of CaMKKβ inhibitor on L-theanine-facilitated phosphorylation of CaMKKβ and AMPKα in HepG2 hepatocytes. HepG2 cells were pretreated with 10 μM BAPTA-AM for 1 h, then added L-theanine co-incubated for 2 h, finally, OA were added co-incubated for 24 h. C Intracellular Ca2+ levels were assessed in HepG2 cells with Fluo-4 AM using a fluorescence microscope. Scale bar: 20 μm (20 ×). D [Ca2+]i was detected by a multifunctional microplate reader using Fluo-4 AM. under the condition of 488 nm excitation wavelength and 520 nm emission wavelength. E Western blots of indicated proteins in HepG2 cells treated with an intracellular Ca2+ chelator BAPTA-AM. Band intensity was quantified by densitometry analysis. Values are expressed as mean ± SEM of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, versus control group; #p < 0.05, ##p < 0.01, versus OA group; &p < 0.05 versus OA + STO-609 group, $p < 0.05 versus OA + BAPTA-AM group. n.s.: not significant (p > 0.05) versus control group
5). Metformin protects against ethanol-induced liver triglyceride accumulation by the LKB1/AMPK/ACC pathway. Molecular Biology Reports (PubMed: 35733070) [IF=2.8]
6). Cholinergic receptors play a role in the cardioprotective effects of anesthetic preconditioning: Roles of nitric oxide and the CaMKKβ/AMPK pathway. Experimental and Therapeutic Medicine (PubMed: 33456504) [IF=2.7]

Application: WB    Species: Rat    Sample: myocardial tissue

Figure 5 Western blot analysis of AMPK, CAMKKβ, AMPK phosphorylation at Thr-172 and CAMKKβ phosphorylation at Ser-511 in the homogenates of myocardial tissue from rat hearts. (A) Western blot bands of AMPK and p-AMPK(Thr172) protein and the ratio of p-AMPK(Thr172)/total AMPK. (B) Western blot bands of CAMKKβ and p-CAMKKβ(Ser511) protein and the ratio of p-CAMKKβ(Ser511)/total CAMKKβ. APC-induced increases of AMPK and CAMKKβ phosphorylation were reduced by the muscarinic acetylcholine receptor antagonist atropine (ATR, 100 nM) and nicotinic acetylcholine receptor antagonist hexamethonium (HEM, 50 µM). The data are presented as the mean ± standard deviation. n=5 hearts/group. *P<0.05 vs. Sham group; &P<0.05 vs. IR group; #P<0.05 vs. APC group. CAMKKβ, calcium/calmodulin-dependent protein kinase kinase β; p, phosphorylated; IR, ischemia-reperfusion; APC, anesthetic preconditioning; ATR, atropine; HEM, hexamethonium; CTL, control.

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