13-9800

antibody from Invitrogen Antibodies

Targeting: CAMK2B CAM2, CAMK2, CAMKB, CaMKIIβ

Provider product page for 13-9800

Western blot

ELISA

Immunoprecipitation

Immunohistochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [29]
Comments [0]
Validations
Flow cytometry [1]
Other assay [17]

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Product number: 13-9800 - Provider product page
Provider: Invitrogen Antibodies
Product name: CaMKII beta Monoclonal Antibody (CB-beta-1)
Antibody type: Monoclonal
Antigen: Purifed from natural sources
Reactivity: Human, Mouse, Rat
Host: Mouse
Isotype: IgG
Antibody clone number: CB-beta-1
Vial size: 100 µg
Concentration: 0.5 mg/mL
Storage: -20°C

CAMK2B protein structure - 13-9800 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 DGLalpha interacts with CaMKII (a) Western blot confirms that CaMKIIalpha specifically associates with DGLalpha immune complexes, but not control IgG complexes, isolated from mouse striatal extracts. (b) CaMKIIalpha was detected in DGLalpha-V5 but not DGLbeta-V5 immune complexes isolated from co-transfected HEK293T cells. Control cells were not transfected or expressed either CaMKIIalpha or DGLalpha/beta-V5 alone. (c) Both CaMKII-alpha and -beta associated with DGLalpha-V5 immune complexes isolated from co-transfected HEK293T cells. The band between CaMKII-alpha and -beta in all of the IP samples is the IgG heavy chain used for the immunoprecipitation, as indicated. Panels a-c are representative of at least two experiments. (d) Alignment of DGLalpha and DGLbeta highlights a unique C-terminal domain in DGLalpha. A GST fusion protein containing the C-terminal domain (CTD; residues 761-1042) of DGLalpha directly interacts with purified Thr 286 -autophosphorylated, but not non-phosphorylated, CaMKIIalpha in vitro . The amount of CaMKII detected in each sample was normalized to the amount detected in an aliquot of the input for each incubation (Non-P (n=5 independent experiments): 0.15+-0.09 and 0.11+-0.03 for GST and GST-DGLalpha-CTD, respectively. P-T286 (n=6 independent experiments): 0.07+-0.03 and 0.37+-0.07 for GST and GST-DGLalpha-CTD, respectively. 2-way ANOVA: Interaction, F(1,18)=7.739, P=0.0123. Autophosphorylation, F 1,18 =4.661, P=0.0446. Protein, F 1,18 =2.22

Supportive validation

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Experimental details: Fig 2 Activation of CaMKII pathway in H9C2 cardiomyoblasts. To investigate the role of the cross talk between CaMKII and ERK on cardiac hypertrophy, we assessed the effect of CaMKII inhibition on PE-induced ERK activation. H9C2s were pretreated with KN93 (5 mumol/L) or AntCaNtide (10 mumol/L) and tat-CN17beta (10 mumol/L) for 30 min and then stimulated with PE (100 nmol/L for 15 min.). A: H9C2 total cell lysates were analyzed by Western blotting (WB) for phosphothreonine 286 CaMKII (pCaMKII) and total CaMKII (CaMKII), with specific antibodies. Data from immunoblots were quantified by densitometric analysis (DA). pCaMKII levels were corrected by total CaMKII densitometry. * = p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 4 PKA inhibition does not modify PE-induced CaMKII/ERK activation in H9C2 cardiomyoblasts. A: H9C2 cardiomyoblasts were transfected with PKA-I and then stimulated with 100 nm phenylephrine (PE) for 15 min. Total cell extracts were analyzed by western blot for phosphothreonine 286 CaMKII (pCaMKII) and CaMKII with specific antibodies. pCaMKII levels were corrected by CaMKII densitometry. *, P < 0.05 vs . Ctrl. B: H9C2s were stimulated with PE after transfection with PKA-I. Total lysates were analysed by WB for pERK with specific antibody. pERK1/2 levels were corrected by ERK1/2 densitometry. * = p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 5 CaMKII/ERK-dependent regulation of the hypertrophy marker ANF. A: H9C2 cells at 70% confluence were incubated 1 h at 37degC with 5 mL DMEM containing purified adenovirus at a multiplicity of infection (moi) of 100:1, encoding either the kinase-dead (CaMKII-DN, rCaMKIIdelta, K42M), or the wild type (CaMKII-WT, rCaMKIIdelta) variant of CaMKII or the empty virus as a negative control (Ctr). 48 h after the infection, the cells were stimulated with PE 100 nM for 24 h. Total RNA was isolated from H9C2s using TRIzol reagent, and cDNA was synthesized by means of a Thermo-Script RT-PCR System, following the manufacturer's instruction. Then ANP gene expression was evaluated by real-time PCR. Results are expressed as mean+-SEM from 3 independent experiments. The ratio of fold change was calculated using the Pfaffl method[ 36 ]. * = p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 1 CaMKII isoforms. A: To evaluate the expression of CaMKII isoforms in ventricular adult cardiomyocytes from WKY rats, the total cell lysates were immunoprecipitated using anti-CaMKII antibody (lane a) or anti-CaMKII alpha, beta gamma or delta specific antibodies (lane b) and together to total samples from WKY cardiomyocytes (lane d) and H9C2 cardiomyoblasts (lane e) were analyzed by western blot with anti-CaMKII alpha, beta gamma or delta antibodies as indicated. Total extracts from rat brain (lane f), mouse brain (lane g) and mouse heart (lane h) were used as controls. Total lysates from WKY cardiomyocytes with specific antibody without A/G agarose beads were used as negative control (lane c). B: Total RNA from H9C2 cell line and isolated ventricular cardiomyocytes was extracted with standard methods. RT-PCR for CaMKII alpha, beta, gamma, and delta was performed as indicated in methods. The representative graph indicates the relative amounts of transcripts for CaMKII isoforms in H9C2s and ventricular adult myocytes from WKY rats. Cycle threshold (Ct) values from 3 independent experiments were normalized to the internal beta-actin control. The ratio of fold change was calculated using the Pfaffl method. * = p

Supportive validation

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Experimental details: Figure 2. Representative immunoblots showing the CaMKIIalpha, CaMKIIbeta and phospho-T286-CaMKIIalpha levels in the hippocampus and amygdala of the kinase-dead CaMKIIalpha (K42R)-KI mouse as compared to the wild-type mouse. The amounts of protein used were 2, 4, and 8 mug for the detection of CaMKIIalpha, CaMKIIbeta, and phospho-T286-CaMKIIalpha (P-CaMKIIalpha), respectively, from hippocampal or amygdala homogenates. Autoradiography of duplicated samples from a pair of wild-type (WT) and kinase-dead CaMKIIalpha (K42R)-KI mice in the same experimental group used for quantitative immunoblot analyses are shown. See also Table 1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 CaMKIIbeta inhibition in the vCA1 increases stress susceptibility (A) AAV vectors used for the control construct (AAV-shControl) and CaMKIIbeta knockdown (AAV-shCaMKIIbeta). ITR, inverted terminal repeats; P CMV , cytomegalovirus promoter; P Camk2a , Camk2a promoter; U6, U6 promoter; WPRE, woodchuck hepatitis virus posttranscriptional regulatory element. (B) Experimental paradigm for behavioral testing. CUMS, chronic ultra-mild stress; SIT, social interaction test; SPT, sucrose preference test; FST, forced swim test. (C) AAV microinjection into the vCA1. Region-specific expression of GFP in vCA1 is shown. Scale bar, 100 mum. (D) Specific knockdown of CaMKIIbeta but not CaMKIIalpha by the AAV-shCaMKIIbeta. (E-G) CaMKIIbeta knockdown in the vCA1 of B6 mice leads to significantly decreased social interaction time in the SIT (E), decreased sucrose preference in SPT (F), and increased immobility time in FST (G) following CUMS exposure. n = 12-15 mice per group. (H) AAV vectors engineered to overexpress a constitutively active form of CaMKIIbeta (CaMKIIbeta-CA) with resistance against shCaMKIIbeta (AAV-CaMKIIbeta-CA Res ) or a control construct (AAV-mCherry). (I) Experimental paradigm for behavioral testing. (J) AAV microinjection into the vCA1. Region-specific expression of GFP and mCherry in the vCA1 is shown. Scale bar, 100 mum. (K and L) Overexpression of CaMKIIbeta-CA Res together with shCaMKIIbeta increases time in the interaction zone in the SIT (K) and tends to inc