Overview

  • Product name

    Anti-BMX (phospho Y566) antibody
    See all BMX primary antibodies
  • Description

    Rabbit polyclonal to BMX (phospho Y566)
  • Host species

    Rabbit
  • Specificity

    This antibody detects endogenous levels of ETK only when phosphorylated at tyrosine 566.
  • Tested applications

    Suitable for: IHC-P, ELISA, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic phosphopeptide derived from human BMX around the phosphorylation site of tyrosine 566 (DQYPVS).

  • Positive control

    • Human thyroid gland tissue

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride

    Without Mg+2 and Ca+2
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Affinity purified from rabbit antiserum by affinity chromatography using epitope specific phosphopeptide. The antibody against non phosphopeptide was removed by chromatography using non phosphopeptide corresponding to the phosphorylation site.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab59409 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use at an assay dependent concentration.
ELISA 1/10000.
WB 1/3000. Detects a band of approximately 74 kDa (predicted molecular weight: 78 kDa).

Target

  • Function

    Activity is required for interleukin 6 (IL-6) induced differentiation. May play a role in the growth and differentiation of hematopoietic cells. May be involved in signal transduction in endocardial and arterial endothelial cells.
  • Tissue specificity

    Preferentially expressed in epithelial and endothelial cells.
  • Sequence similarities

    Belongs to the protein kinase superfamily. Tyr protein kinase family. TEC subfamily.
    Contains 1 Btk-type zinc finger.
    Contains 1 PH domain.
    Contains 1 protein kinase domain.
    Contains 1 SH2 domain.
  • Domain

    SH2 domain mediates interaction with RUFY1.
  • Cellular localization

    Cytoplasm.
  • Information by UniProt
  • Database links

  • Alternative names

    • Bmx antibody
    • BMX non receptor tyrosine kinase antibody
    • BMX_HUMAN antibody
    • BONE MARROW KINASE, X-LINKED antibody
    • Bone marrow tyrosine kinase gene in chromosome X protein antibody
    • Cytoplasmic tyrosine protein kinase BMX antibody
    • Cytoplasmic tyrosine-protein kinase BMX antibody
    • Epithelial and endothelial tyrosine kinase antibody
    • ETK antibody
    • NKT38 antibody
    • NTK 38 antibody
    • NTK38 antibody
    • Protein tyrosine kinase BMX antibody
    • PSCTK 2 antibody
    • PSCTK 3 antibody
    • PSCTK2 antibody
    • PSCTK3 antibody
    see all

Images

  • Immunohistochemical analysis of paraffin embedded human thyroid gland tissue using ab59409 at a 1/50 dilution, in the presence (right) and absence (left) of immunizing phosphopeptide.
  • Anti-BMX (phospho Y566) antibody (ab59409) at 1 µg/ml + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Predicted band size: 78 kDa
    Observed band size: 74 kDa
    why is the actual band size different from the predicted?

  • All lanes : Anti-BMX (phospho Y566) antibody (ab59409)

    Lane 1 : Extracts from HeLa cells treated with Serum 20% 15'
    Lane 2 : Extracts from HeLa cells treated with Serum 20% 15' and with the synthesized peptide.

    Predicted band size: 78 kDa

References

This product has been referenced in:

  • Holopainen T  et al. Endothelial Bmx tyrosine kinase activity is essential for myocardial hypertrophy and remodeling. Proc Natl Acad Sci U S A 112:13063-8 (2015). Read more (PubMed: 26430242) »
  • Guryanova OA  et al. Nonreceptor Tyrosine Kinase BMX Maintains Self-Renewal and Tumorigenic Potential of Glioblastoma Stem Cells by Activating STAT3. Cancer Cell 19:498-511 (2011). WB, ICC/IF ; Mouse . Read more (PubMed: 21481791) »
See all 2 Publications for this product

Customer reviews and Q&As

Question
Answer

Bitte entschuldigen Sie die Verzögerung.

Ich habe nun aus dem Labor die Protokolle sowohl für die Behandlung der HeLa Zellen als auch das WB Protokoll für ab59409 bekommen.

Ich kopiere ihnen beide Protokolle unten in die Email.

Bitte lassen Sie mich wissen ob die Protokolle geholfen haben.

The protocol of sample treatment for Western blot (for ab59409) is following:

Grow Hela cells to ˜80% confluence in DMEM with 10% FBS at 37C and 5% CO2

Remove the culture medium and wash with DMEM without FBS

Starve the cells in DMEM with 0.2% FBS for 72 hours at 37C and 5% CO2 with changes starving medium (DMEM with 0.2% FBS) once a day (total three times)

After 72 hours, remove the medium, culture the cells in DMEM medium with 20% FBS (fresh) for 15 minutes at 37C and 5% CO2

Remove the medium and rinse the cells with 1x PBS buffer two times

Lyse the cells in lysis buffer and measure protein concentration

Run 10% SDS-PAGE with 50 ug per lane for 150V (for mini-gel)

Transfer to NC paper for Western blot





Blocking

1. Block membrane by incubating 1 hour at room temperature or overnight at 4℃ with shaking in Blocking Solution (5% BSA, 0.05% Tween-20 in TBS (50mM Tris, 100mM NaCl, pH 7.6).

Incubation with Primary Antibody

2. Dilute primary antibody at the appropriate dilution in Blocking Solution.

3. Incubate the membrane with diluted primary antibody overnight at 4℃ with agitation.

4. Remove antibody solution. Wash the membrane 3 times for 5-10 minutes each time at room temperature in TBST (50mM Tris, 100mM NaCl, 0.05% Tween-20, pH 7.6) with shaking. Note: Increase the concentration of Tween-20 to 0.1% reduces the background and increases the specificity, but it will reduce the sensitivity.

Incubation with Second Antibody

5. Incubate membrane with secondary AP conjugate diluted (according to manufacturer’s instructions) in Blocking Solution for 1 hour at room temperature with shaking.

6. Repeat Step 4.

7. Wash membrane with TBS for 2-5 minutes before proceeding Chemiluminescent Reaction.

Chemiluminescent Reaction

8. Prepare and use the Chemiluminescent substrate according to the manufacturer’s instructions.

9. Immediately wrap the membrane and expose to X-ray films for 10 second to 1 hour period. The exposure time may vary according to the amount of antibody and antigen.

Read More

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