Invitrogen

CaMKI Polyclonal Antibody

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

View all (6) CaMKI antibodies

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Datasheet

Protocols

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CaMKI Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of CaM Kinase I was performed using 70% confluent log phase U-87MG cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with CaM Kinase I Rabbit Polyclonal Antibody (Product # PA1-543) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

CaMKI Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of CaM Kinase I showing staining in the cytoplasm and nucleus of paraffin-embedded mouse kidney tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a CaM Kinase I Rabbit Polyclonal Antibody (Product # PA1-543) diluted in 3% BSA-PBS at a dilution of 1:20 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

CaMKI Antibody in Western Blot (WB) — Western blot analysis was performed on tissue extracts (30 µg lysate) of Rat Brain (Lane 1), and Mouse Brain (Lane 2). The blots were probed with Anti-CaM Kinase I Rabbit Polyclonal Antibody (Product # PA1-543, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A ~ 41 kDa band corresponding to CaM Kinase I was observed across tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

CaMKI Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across tissue models owing to their inherent genetic constitution. Relative expression of CaMKI was observed in Mouse Brain, Rat Brain, Rat Liver and Rat Kidney in Western Blot using CaMKI Polyclonal Antibody (Product # PA1-543). CaMKI is reported to be expressed in brain tissue and not in other tissues like liver and kidney. {RE}

Product Details

PA1-543

Applications	Tested Dilution	Publications
Western Blot (WB)	1 µg/mL	-
Immunohistochemistry (Paraffin) (IHC (P))	1:20-1:100	-
Immunocytochemistry (ICC/IF)	2 µg/mL	-

Product Specifications
Species Reactivity	Human, Mouse, Rat
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Synthetic Peptide: A(1) V E G P R W K Q A E D I R D I Y D(18) View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS with 1mg/mL BSA
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2070105

Product Specific Information

PA1-543 detects calciµm/calmodulin-dependent kinase I (CaMKI) from rat and mouse brain extract.

PA1-543 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~43 kDa protein representing CaMKI from rat and mouse brain extract.

PA1-543 immunizing peptide corresponds to amino acid residues 1-18 from human CaMKI protein. PA1-543 immunizing peptide (Cat. # PEP-134) is available for use in neutralization and control experiments.

Target Information

Calcium/calmodulin-dependent protein kinase I is expressed in many tissues and is a component of a calmodulin-dependent protein kinase cascade. Calcium/calmodulin directly activates calcium/calmodulin-dependent protein kinase I by binding to the enzyme and indirectly promotes the phosphorylation and synergistic activation of the enzyme by calcium/calmodulin-dependent protein kinase I kinase.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: calcium/calmodulin-dependent protein kinase I; Calcium/calmodulin-dependent protein kinase type 1; CaM kinase I; CaM kinase I alpha; CaM-KI; CAMK1-PEN; CaMKI-alpha; G alpha interacting protein; MGC120317; MGC120318; regulator of G-protein signalling 19

Gene Aliases: AI505105; CAMK1; CAMKI; CaMKIalpha; D6Ertd263e; Gaip

UniProt ID: (Human) Q14012, (Rat) Q63450, (Mouse) Q91YS8

Entrez Gene ID: (Human) 8536, (Rat) 171503, (Mouse) 52163

Molecular Function: non-receptor serine/threonine protein kinase