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Creatine kinase B type Recombinant Rabbit Monoclonal Antibody [JB78-34]

Usd: 385

Specification

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Catalog# ET7107-52

Creatine kinase B type Recombinant Rabbit Monoclonal Antibody [JB78-34]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Creatine kinase B type Recombinant Rabbit Monoclonal Antibody [JB78-34]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human Creatine kinase B type aa 27-118 / 381.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, FC

Molecular Weight

Predicted band size: 43 kDa

Positive Control

HeLa cell lysate, SH-SY5Y cell lysate, Y79 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, HAP1 cell lysate, mouse brain tissue, 293T, LOVO.

Conjugation

unconjugated

Clone Number

JB78-34

RRID

AB_3073480

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:50-1:200

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:1,000

  • FC

  • 1:50-1:100

Target

Function

Brain-type creatine kinase also known as CK-BB is a creatine kinase that in humans is encoded by the CKB gene. The protein encoded by this gene, CK-BB, consists of a homodimer of two identical brain-type CK-B subunits. BB-CK is a cytoplasmic enzyme involved in cellular energy homeostasis, with certain fractions of the enzyme being bound to cell membranes, ATPases, and a variety of ATP-requiring enzymes in the cell. There, CK-BB forms tightly coupled microcompartments for in situ regeneration of ATP that has been used up. The encoded protein reversibly catalyzes the transfer of "energy-rich" phosphate between ATP and creatine or between phospho-creatine (PCr) and ADP. Its functional entity is a homodimer (CK-BB) in brain and smooth muscle as well as in other tissues and cells such as neuronal cells, retina, kidney, bone, etc. In heart, a heterodimer (CK-MB) shahil consisting of one CK-B brain-type CK subunit and one CK-M muscle-type CK subunit is prominently expressed. The encoded CK-BB and CK-MB proteins are members of the ATP:guanido phosphotransferase protein family.

Background References

1. Bong S M et al. Structural studies of human brain-type creatine kinase complexed with the ADP-Mg2+-NO3- -creatine transition-state analogue complex. FEBS Lett 582:3959-3965 (2008).

2. Daouk G H et al. Isolation of a functional human gene for brain creatine kinase. J Biol Chem 263:2442-2446 (1988).

Sequence Similarity

Belongs to the ATP:guanido phosphotransferase family.

Subcellular Location

Cytoplasm, cytosol, Mitochondrion, Cell membrane.

UNIPROT

SwissProt: P12277 Human

SwissProt: Q04447 Mouse

SwissProt: P07335 Rat

Synonyms

B CK antibody

B-CK antibody

BB-CK antibody

BCK antibody

Brain creatine kinase antibody

Ckb antibody

CKBB antibody

Creatine kinase B antibody

Creatine kinase B chain antibody

Creatine kinase B type antibody

Expand

Images

  • Western blot analysis of Creatine kinase B type on different lysates with Rabbit anti-Creatine kinase B type antibody (<a href="/products/ET7107-52" style="font-weight: bold;text-decoration: underline;">ET7107-52</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: SH-SY5Y cell lysate<br />Lane 3: Y79 cell lysate<br />Lane 4: Mouse brain tissue lysate<br />Lane 5: Rat brain tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 43 kDa<br />Observed band size: 43 kDa<br /><br />Exposure time: 4 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7107-52" style="font-weight: bold;text-decoration: underline;">ET7107-52</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Creatine kinase B type on different lysates with Rabbit anti-Creatine kinase B type antibody (ET7107-52) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: SH-SY5Y cell lysate
    Lane 3: Y79 cell lysate
    Lane 4: Mouse brain tissue lysate
    Lane 5: Rat brain tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 43 kDa
    Observed band size: 43 kDa

    Exposure time: 4 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7107-52) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Creatine kinase B type on different lysates with Rabbit anti-Creatine kinase B type antibody (<a href="/products/ET7107-52" style="font-weight: bold;text-decoration: underline;">ET7107-52</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Creatine kinase B type KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 43 kDa<br />Observed band size: 43 kDa<br /><br />Exposure time: 6 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7107-52" style="font-weight: bold;text-decoration: underline;">ET7107-52</a>) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of Creatine kinase B type on different lysates with Rabbit anti-Creatine kinase B type antibody (ET7107-52) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Creatine kinase B type KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 43 kDa
    Observed band size: 43 kDa

    Exposure time: 6 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7107-52) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Creatine kinase B type antibody (<a href="/products/ET7107-52" style="font-weight: bold;text-decoration: underline;">ET7107-52</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7107-52" style="font-weight: bold;text-decoration: underline;">ET7107-52</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Creatine kinase B type antibody (ET7107-52) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7107-52) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • ICC staining of Creatine kinase B type in 293T cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET7107-52" style="font-weight: bold;text-decoration: underline;">ET7107-52</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of Creatine kinase B type in 293T cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7107-52, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of Creatine kinase B type in LOVO cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET7107-52" style="font-weight: bold;text-decoration: underline;">ET7107-52</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of Creatine kinase B type in LOVO cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7107-52, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • Flow cytometric analysis of Creatine kinase B type was done on 293T cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET7107-52" style="font-weight: bold;text-decoration: underline;">ET7107-52</a>, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of Creatine kinase B type was done on 293T cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7107-52, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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