Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23430-12] to HEXIM1 - ChIP Grade
  • Suitable for: WB, ICC/IF, IP, IHC-P, ChIP, Flow Cyt
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade
    See all HEXIM1 primary antibodies

  • Description

    Rabbit monoclonal [EPR23430-12] to HEXIM1 - ChIP Grade

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Mouse
    Human
    Flow Cyt
    Human
    ICC/IF
    Mouse
    Human
    IHC-P
    Human
    IP
    Mouse
    Human
    WB
    Rat
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: LNCaP, PC-12 whole cell lysates. IHC-P: Human cardiac muscle and testis tissue. ICC/IF: HeLa and MEF cells. Flow Cyt: HeLa cells. IP: HeLa (treated with 10 M MG-132 for 24 hours) and MEF whole cell lysates.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Images

  • Western blot - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)
    Western blot - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)
    All lanes : Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647) at 1/1000 dilution

    Lane 1 : LNCaP (human prostate carcinoma epithelial cell) whole cell lysate
    Lane 2 : PC-12 (rat adrenal gland pheochromocytoma ) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051))
    Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 41 kDa
    Observed band size: 60 kDa
    why is the actual band size different from the predicted?



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

    The molecular weight observed is consistent with what has been described in the literature (PMID: 28254838)

    Lysates were made freshly and used in WB test immediately to minimize protein degradation. 

    Exposure time: 26 seconds.

  • Immunoprecipitation - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)
    Immunoprecipitation - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)

    HEXIM1 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) (treated with 10 μM MG-132 for 24 hours), whole cell lysate with ab240647 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab240647 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) treated with 10 μM MG-132 for 24 hours, whole cell lysate 10 ug

    Lane 2: ab240647 IP in HeLa treated with 10 μM MG-132 for 24 hours whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab240647 in HeLa treated with 10 μM MG-132 for 24 hours whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

     

  • Immunoprecipitation - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)
    Immunoprecipitation - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)

    HEXIM1 was immunoprecipitated from 0.35 mg MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate with ab240647 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab240647 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate 10 ug

    Lane 2: ab240647 IP in MEF whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab240647 in MEF whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    Lysate were made freshly and used in IP test immediately to minimize protein degradation. Incubation for immunoprecipitation was carried out overnight at 4°C.

  • ChIP - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)
    ChIP - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)

    Chromatin was prepared from MEF cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab 240647 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
    Primers and probes are from paper PMCID: PMC4103662

    *https://www.abcam.com/resources?keywords=X%20ChIP%20protocol

  • Immunocytochemistry/ Immunofluorescence - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)
    Immunocytochemistry/ Immunofluorescence - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)

    Immunofluorescent analysis of 100% methanol-fixed MEF cells labelling HEXIM1 with ab240647 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000  (Green). Confocal image showing nuclear staining in MEFs. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000  dilution.

  • Flow Cytometry - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)
    Flow Cytometry - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)

    Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling HEXIM1 with ab240647 at 1/500 dilution (0.1ug) (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, ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)

    Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling HEXIM1 with ab240647 at 1/2000 dilution  dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human testis (PMID: 23300697). The section was incubated with ab240647 for 10 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

  • ChIP - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)
    ChIP - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)

    Chromatin was prepared from LNCaP cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
    The ChIP was performed with 25 µg of chromatin, 5 µg of ab240647 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 20 µl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).

    https://www.abcam.com/resources?keywords=X%20ChIP%20protocol

  • Immunocytochemistry/ Immunofluorescence - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)
    Immunocytochemistry/ Immunofluorescence - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)

    Immunofluorescent analysis of 100% methanol-fixed HeLa cells labelling HEXIM1 with ab240647 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000  (Green). Confocal image showing nuclear staining in HeLa cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200  dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)

    Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling HEXIM1 with ab240647 at 1/2000 dilution dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human cardiac muscle (PMID: 23300697). The section was incubated with ab240647 for 10 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

  • Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)
    Anti-HEXIM1 antibody [EPR23430-12] - ChIP Grade (ab240647)

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