Anti-Creatine Kinase MM antibody [EPR25358-10]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Creatine Kinase MM antibody [EPR25358-10] (AB283683)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A-673 cells labelling Creatine Kinase MM with ab283683 at 1/100 (5.39 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in A-673 cell line. is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Creatine Kinase MM antibody [EPR25358-10] (AB283683)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labelling Creatine Kinase MM with ab283683 at 1/10000 (0.054 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Negative control : No staining on the human spleen. The section was incubated with ab283683 at 4 ° overnight. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Creatine Kinase MM antibody [EPR25358-10] (AB283683)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labelling Creatine Kinase MM with ab283683 at 1/5000 (0.108 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human skeletal muscle. The section was incubated with ab283683 at 4° overnight. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• mIHC

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Multiplex immunohistochemistry - Anti-Creatine Kinase MM antibody [EPR25358-10] (AB283683)

Fluorescence multiplex immunohistochemical analysis of the human skeletal muscle (Formalin/PFA-fixed paraffin-embedded sections). Panel A : merged staining of anti-Dystrophin (ab275391, red; Opal™690), anti-A2BP1/Fox1/RBFOX1 (ab254413, green; Opal™520) and anti-Creatine Kinase MM (ab283683, magenta; Opal™570) on human skeletal muscle. Panel B : anti-A2BP1/Fox1/RBFOX1 stained on nucleus of skeletal muscle. Panel C : anti-Creatine Kinase MM stained on cytoplasm of skeletal muscle. Panel D : anti-Dystrophin stained on membrane of skeletal muscle. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The section was incubated in three rounds of staining : in the order of ab275391 at 1/500 (2.08 μg/ml) dilution, ab254413 at 1/2000 (0.239 μg/ml) dilution, and ab283683 at 1/3000 (0.18 μg/ml) dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. DAPI (blue) was used as a nuclear counter stain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Creatine Kinase MM antibody [EPR25358-10] (AB283683)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labelling Creatine Kinase MM with ab283683 at 1/10000 (0.054 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Cytoplasmic staining on mouse cardiac muscle. The section was incubated with ab283683 at 4° overnight. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-Creatine Kinase MM antibody [EPR25358-10] (AB283683)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse spleen (fresh) tissue labeling Creatine Kinase MM with ab283683 at 1/100 (5.39 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green). No staining on mouse spleen is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Creatine Kinase MM antibody [EPR25358-10] (AB283683)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cardiac muscle (fresh) tissue labeling Creatine Kinase MM with ab283683 at 1/100 (5.39 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green). Cytoplasmic staining on mouse cardiac muscle is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Creatine Kinase MM antibody [EPR25358-10] (AB283683)

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labelling Creatine Kinase MM with ab283683 at 1/10000 (0.054 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on rat cardiac muscle. The section was incubated with ab283683 at 4° overnight. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Creatine Kinase MM antibody [EPR25358-10] (AB283683)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cardiac muscle (fresh) tissue labeling Creatine Kinase MM with ab283683 at 1/100 (5.39 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green). Cytoplasmic staining on rat cardiac muscle is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Creatine Kinase MM antibody [EPR25358-10] (AB283683)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat spleen (fresh) tissue labeling Creatine Kinase MM with ab283683 at 1/100 (5.39 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (Green). No staining on rat spleen is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Creatine Kinase MM antibody [EPR25358-10] (AB283683)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A-673 (Human muscle Ewing's Sarcoma) cells labelling Creatine Kinase MM with ab283683 at 1/50 dilution (1ug)/ red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• WB

Lab

Western blot - Anti-Creatine Kinase MM antibody [EPR25358-10] (AB283683)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 1 second

All lanes:

Western blot - Anti-Creatine Kinase MM antibody [EPR25358-10] (ab283683) at 1/2000 dilution

All lanes:

Human heart tissue lysate

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 43 kDa

false

• WB

Lab

Western blot - Anti-Creatine Kinase MM antibody [EPR25358-10] (AB283683)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTCK protein level is low in undifferentiated myoblasts and becomes strikingly upregulated as differentiation progresses. The expression profile observed is consistent with what has been described in the literature (PMID : 22703875).

Exposure time : 3 minutes

All lanes:

Western blot - Anti-Creatine Kinase MM antibody [EPR25358-10] (ab283683) at 1/1000 dilution

Lane 1:

Untreated C2C12 (mouse myoblasts myoblast) whole cell lysate

Lane 2:

C2C12 treated with 2% horse serum for 6 days whole cell lysate

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 43 kDa

false

• WB

Lab

Western blot - Anti-Creatine Kinase MM antibody [EPR25358-10] (AB283683)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : Lanes 1-2 : 3 seconds, Lane 3 : 48 seconds; Lanes 4-5 : 6 seconds.

The molecular weight observed is consistent with what has been described in the literature (PMID : 22066028).

No expression was observed in spleen.

All lanes:

Western blot - Anti-Creatine Kinase MM antibody [EPR25358-10] (ab283683) at 1/2000 dilution

Lane 1:

Mouse heart tissue lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Lane 3:

A-673 (Human muscle Ewing's sarcoma) whole cell lysate at 20 µg

Lane 4:

Rat heart tissue lysate at 20 µg

Lane 5:

Rat spleen tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 43 kDa

Observed band size: 43 kDa

false