Anti-CaMKII (Alpha-Specific) Monoclonal Antibody

Product Name

Anti-CaMKII (Alpha-Specific) Monoclonal Antibody

View all Camk2a Antibodies

SKU/Catalog Number

M03241-1

Size

100ug

Form

liquid

Description

Boster Bio Anti-CaMKII (Alpha-Specific) Monoclonal Antibody catalog # M03241-1. Tested in ELISA, IP, IF, IHC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20°C for one year. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-CaMKII (Alpha-Specific) Monoclonal Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # M03241-1)

Host

Mouse

Contents

PBS pH7.4, 50% glycerol, 0.09% sodium azide

Clonality

Monoclonal

Clone Number

6G9

Isotype

IgG1

Immunogen

Partially purified rat CaMKII

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

Detects ~50-60kDa. Recognizes both phosphorylated and non-phosphorylated forms.

Reactive Species

M03241-1 is reactive to Camk2a in Bovine, Human, Mouse, Rat

Applications

M03241-1 is guaranteed for ELISA, IP, IF, IHC, ICC, WB, RIA Boster Guarantee

Observed Molecular Weight

42 kDa

Calculated molecular weight

10659 MW

Background of Camk2a

CaMKII is an important member of the calcium/calmodulin-activated protein kinase family, functioning in neural synaptic stimulation and T-cell receptor signaling (1, 2). CaMKII is expressed in many different tissues but is specifically found in the neurons of the forebrain and its mRNA is found within the dendrites and the soma of the neuron. The CaMKII that is found in the neurons consist of two subunits of 52 (termed alpha genes) and 60 kDa (beta genes). CaMKII has catalytic and regulatory domains, as well as an ATP-binding domain, and a consensus phosphorylation site (3-7). The binding of Ca2+/calmodulin to its regulatory domain releases its auto inhibitory effect and activates the kinase (8). This kinase activation results in autophosphorylation at threonine 286 (8). The threonine phosphorylation state of CaMKII can be regulated through PP1/PKA. Whereas PP1 (protein phosphatase 1) dephosphorylates phospho-CaMKII at Thr286, PKA (protein kinase A) prevents this dephosphorylation (9). Autophosphorylation also enables CaMKII to attain an enhanced affinity for NMDA receptors in postsynaptic densities (10-12).

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

WB (1:10000), IHC (1:2000), ICC/IF (1:50); optimal dilutions for assays should be determined by the user.

Validation Images & Assay Conditions

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Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-CaMKII Monoclonal Antibody, Clone 6G9 (M03241-1) . Tissue: dissociated hippocampal neurons. Species: Mouse. Fixation: Cold 4% paraformaldehyde/0.2% glutaraldehyde in 0.1M sodium phosphate buffer. Primary Antibody: Mouse Anti-CaMKII Monoclonal Antibody (M03241-1) at 1:1000 for 12 hours at 4°C. Secondary Antibody: FITC Goat Anti-Mouse IgG (green) at 1:50 for 30 minutes at RT. Magnification: 10X. Courtesy of: Mary Kennedy, Caltech.

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Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-CaMKII Monoclonal Antibody, Clone 6G9 (M03241-1) . Tissue: HaCaT cells. Species: Human. Fixation: Cold 100% methanol for 10 minutes at -20°C. Primary Antibody: Mouse Anti-CaMKII Monoclonal Antibody (M03241-1) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Localization: Nuclear Staining.

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Figure 2. Western blot analysis of Camk2a using anti-Camk2a antibody (M03241-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Camk2a antigen affinity purified polyclonal antibody (Catalog # M03241-1) at 0.5 ug/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-Mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # SA1021) with Tanon 5200 system. A specific band was detected for Camk2a.

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Figure 3. IHC analysis of Camk2a using anti-Camk2a antibody (M03241-1).
Camk2a was detected in paraffin-embedded section. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-Camk2a Antibody (M03241-1) overnight at 4°C. Biotinylated goat anti Mouse IgG antibody was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.

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Figure 4. IHC analysis of Camk2a using anti-Camk2a antibody (M03241-1).
Camk2a was detected in paraffin-embedded section. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-Camk2a Antibody (M03241-1) overnight at 4°C. Biotinylated goat anti Mouse IgG antibody was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.

Gene/Protein Information For Camk2a (Source: Uniprot.org, NCBI)

Alternative Names

calcium/calmodulin-dependent protein kinase (CaM kinase) II alpha; calcium/calmodulin-dependent protein kinase II alpha; calcium/calmodulin-dependent protein kinase II alpha-B subunit; CaM kinase II alpha subunit; CaM Kinase II alpha; CaM kinase II subunit alpha; CAMK2A; CAMKAcalcium/calmodulin-dependent protein kinase type II subunit alpha; CaMK-II alpha subunit; CaMK-II subunit alpha; CaMKIINalpha; CaM-kinase II alpha chain; EC 2.7.11; EC 2.7.11.17; KIAA0968calcium/calmodulin-dependent protein kinase type II alpha chain Camk2a|CaMKII, R74975, mKIAA0968|calcium/calmodulin-dependent protein kinase II alpha|calcium/calmodulin-dependent protein kinase type II subunit alpha|CaMK II|alpha-CaMKII|caM kinase II subunit alpha|caMK-II subunit alpha

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