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Signal Transduction Protein Trafficking Chaperones Heat Shock Proteins
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RecombinantRabMAb

Recombinant Anti-STUB1/CHIP antibody [EPR4447] (ab134064)

  • Datasheet
  • SDS
Reviews (9) Submit a question References (10)

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Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Immunocytochemistry/ Immunofluorescence - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Immunoprecipitation - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Flow Cytometry (Intracellular) - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
  • Anti-STUB1/CHIP antibody [EPR4447] (ab134064)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4447] to STUB1/CHIP
  • Suitable for: WB, IP, Flow Cyt (Intra), ICC/IF, IHC-P
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

APC Carrier Free HRP PE

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Overview

  • Product name

    Anti-STUB1/CHIP antibody [EPR4447]
    See all STUB1/CHIP primary antibodies
  • Description

    Rabbit monoclonal [EPR4447] to STUB1/CHIP
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, Flow Cyt (Intra), ICC/IF, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human STUB1/CHIP aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • HeLa, MCF-7, 293T and ECV-304 cell lysates; Human skeletal muscle tissue.
  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR4447
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Trafficking
    • Chaperones
    • Heat Shock Proteins
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • SMADs
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Ubiquitin ligases
    • Epigenetics and Nuclear Signaling
    • Ubiquitin & Ubiquitin Like Modifiers
    • E3 Ubiquitin Ligases
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-STUB1/CHIP antibody [EPR4447] - BSA and Azide free (ab238966)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab134064 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
WB (5)
1/10000 - 1/50000. Predicted molecular weight: 35 kDa.
IP (2)
1/120.

For unpurified, use 1/10 - 1/100.

Flow Cyt (Intra)
Use at an assay dependent concentration.
ICC/IF (2)
1/250 - 1/500.
IHC-P
1/100.
Notes
WB
1/10000 - 1/50000. Predicted molecular weight: 35 kDa.
IP
1/120.

For unpurified, use 1/10 - 1/100.

Flow Cyt (Intra)
Use at an assay dependent concentration.
ICC/IF
1/250 - 1/500.
IHC-P
1/100.

Target

  • Function

    E3 ubiquitin-protein ligase which targets misfolded chaperone substrates towards proteasomal degradation. Ubiquitinates NOS1 in concert with Hsp70 and Hsp40. Modulates the activity of several chaperone complexes, including Hsp70, Hsc70 and Hsp90. Mediates transfer of non-canonical short ubiquitin chains to HSPA8 that have no effect on HSPA8 degradation. Mediates polyubiquitination of DNA polymerase beta (POLB) at 'Lys-41', 'Lys-61' and 'Lys-81', thereby playing a role in base-excision repair: catalyzes polyubiquitination by amplifying the HUWE1/ARF-BP1-dependent monoubiquitination and leading to POLB-degradation by the proteasome. Mediates polyubiquitination of CYP3A4.
  • Tissue specificity

    Highly expressed in skeletal muscle, heart, pancreas, brain and placenta. Detected in kidney, liver and lung.
  • Pathway

    Protein modification; protein ubiquitination.
  • Sequence similarities

    Contains 3 TPR repeats.
    Contains 1 U-box domain.
  • Domain

    The TPR domain is essential for ubiquitination mediated by UBE2D1.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
    Auto-ubiquitinated; mediated by UBE2D1 and UBE2D2.
  • Cellular localization

    Cytoplasm.
  • Target information above from: UniProt accession Q9UNE7 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 10273 Human
    • Entrez Gene: 56424 Mouse
    • Entrez Gene: 287155 Rat
    • Omim: 607207 Human
    • SwissProt: Q9UNE7 Human
    • SwissProt: Q9WUD1 Mouse
    • Unigene: 592081 Human
    • Alternative names

      • Antigen NY CO 7 antibody
      • Antigen NY-CO-7 antibody
      • C terminus of Hsp70-interacting protein antibody
      • Carboxy terminus of Hsp70 interacting protein antibody
      • Carboxy terminus of Hsp70-interacting protein antibody
      • Carboxy terminus of Hsp70p interacting protein antibody
      • CHIP antibody
      • CHIP_HUMAN antibody
      • CLL associated antigen KW 8 antibody
      • CLL-associated antigen KW-8 antibody
      • E3 ubiquitin protein ligase CHIP antibody
      • E3 ubiquitin-protein ligase CHIP antibody
      • Heat shock protein A binding protein 2 (c terminal) antibody
      • HSPABP2 antibody
      • NY CO 7 antibody
      • PP1131 antibody
      • SDCCAG7 antibody
      • Serologically defined colon cancer antigen 7 antibody
      • STIP1 homology and U Box containing protein 1 antibody
      • STIP1 homology and U box containing protein 1 E3 ubiquitin protein ligase antibody
      • STIP1 homology and U box-containing protein 1 antibody
      • STUB 1 antibody
      • STUB1 antibody
      • UBOX 1 antibody
      • UBOX1 antibody
      see all

    Images

    • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
      Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
      All lanes : Anti-STUB1/CHIP antibody [EPR4447] (ab134064) at 1/20000 dilution (purified)

      Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
      Lane 2 : MCF7 (Human breast adenocarcinoma cell line) cell lysate
      Lane 3 : HEK293 (Human epithelial cell line from embryonic kidney) cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 35 kDa
      Observed band size: 35 kDa



      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Immunocytochemistry/ Immunofluorescence - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
      Immunocytochemistry/ Immunofluorescence - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)

      Immunofluorescence staining of SH-SY5Y cells with purified ab134064 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab134064 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

    • Immunoprecipitation - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
      Immunoprecipitation - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)

      ab134064 (purified) at 1/120 immunoprecipitating STUB1/CHIP in 10 μg HeLa (Lanes 1 and 2, observed at 35 kDa). Lane 3 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)

      Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labelling STUB1/CHIP with unpurified ab134064 at 1/100 dilution.

    • Flow Cytometry (Intracellular) - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
      Flow Cytometry (Intracellular) - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)

      Intracellular Flow Cytometry analysis of SH-SY5Y (Human epithelial cell line from embryonic kidney) cells labeling STUB1/CHIP with purified ab134064 at 1/120 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. 

       

       

    • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
      Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
      Anti-STUB1/CHIP antibody [EPR4447] (ab134064) at 1/10000 dilution (purified) + Mouse brain at 10 µg

      Secondary
      HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

      Predicted band size: 35 kDa
      Observed band size: 35 kDa



      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
      Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)This image is courtesy of an anonymous Abreview
      All lanes : Anti-STUB1/CHIP antibody [EPR4447] (ab134064) at 1/1000 dilution (unpurified)

      Lane 1 : Wild type primary Mouse neurons whole cell lysate
      Lane 2 : STUB1 knockout primary Mouse neurons whole cell lysate
      Lane 3 : Wild type MEF (Mouse embryonic fibroblast cell line) whole cell lysate
      Lane 4 : STUB1 knockout MEF (Mouse embryonic fibroblast cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/20000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 35 kDa


      Exposure time: 25 seconds

      See Abreview

    • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
      Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
      Anti-STUB1/CHIP antibody [EPR4447] (ab134064) at 1/50000 dilution (purified) + Rat brain at 10 µg

      Secondary
      HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

      Predicted band size: 35 kDa
      Observed band size: 35 kDa



      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
      Western blot - Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
      All lanes : Anti-STUB1/CHIP antibody [EPR4447] (ab134064) at 1/10000 dilution (unpurified)

      Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
      Lane 2 : MCF-7 (Human breast adenocarcinoma cell line) cell lysate
      Lane 3 : 293T cell lysate
      Lane 4 : ECV-304 (Human urinary bladder cancer cell line) cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 35 kDa



      ECV-304 (derived from T24, discontinued by ATCC)

    • Anti-STUB1/CHIP antibody [EPR4447] (ab134064)
      Anti-STUB1/CHIP antibody [EPR4447] (ab134064)

    Protocols

    • Flow cytometry protocols
    • Immunoprecipitation protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (10)

    Publishing research using ab134064? Please let us know so that we can cite the reference in this datasheet.

    ab134064 has been referenced in 10 publications.

    • Liu Y  et al. CHIP-mediated CIB1 ubiquitination regulated epithelial-mesenchymal transition and tumor metastasis in lung adenocarcinoma. Cell Death Differ 28:1026-1040 (2021). PubMed: 33082516
    • Wu HH  et al. Hsp70 acts as a fine-switch that controls E3 ligase CHIP-mediated TAp63 and ?Np63 ubiquitination and degradation. Nucleic Acids Res 49:2740-2758 (2021). PubMed: 33619536
    • Kanamori T  et al. Investigation of energy metabolic dynamism in hyperthermia-resistant ovarian and uterine cancer cells under heat stress. Sci Rep 11:14726 (2021). PubMed: 34282188
    • Chen BH  et al. Hsc70/Stub1 promotes the removal of individual oxidatively stressed peroxisomes. Nat Commun 11:5267 (2020). PubMed: 33077711
    • Schuster S  et al. CHIP mutations affect the heat shock response differently in human fibroblasts and iPSC-derived neurons. Dis Model Mech 13:N/A (2020). PubMed: 33097556
    • Zhang L  et al. Cimetidine promotes STUB1-mediated degradation of tumoral FOXP3 by activating PI3K-Akt pathway in gastric cancer. Ann Transl Med 8:1304 (2020). PubMed: 33209884
    • Dema A  et al. Cyclin-Dependent Kinase 18 Controls Trafficking of Aquaporin-2 and Its Abundance through Ubiquitin Ligase STUB1, Which Functions as an AKAP. Cells 9:N/A (2020). PubMed: 32164329
    • Wang Y  et al. STUB1 is targeted by the SUMO-interacting motif of EBNA1 to maintain Epstein-Barr Virus latency. PLoS Pathog 16:e1008447 (2020). PubMed: 32176739
    • Pan C  et al. Hsp90B enhances MAST1-mediated cisplatin resistance by protecting MAST1 from proteosomal degradation. J Clin Invest 129:4110-4123 (2019). PubMed: 31449053
    • Li H  et al. WWP2 is a physiological ubiquitin ligase for phosphatase and tensin homolog (PTEN) in mice. J Biol Chem 293:8886-8899 (2018). PubMed: 29685889

    Customer reviews and Q&As

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    1-9 of 9 Abreviews or Q&A

    Western blot abreview for Anti-STUB1/CHIP antibody [EPR4447]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Monkey Tissue lysate - whole (Heart)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    30 µg
    Specification
    Heart
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 01 2019

    Western blot abreview for Anti-STUB1/CHIP antibody [EPR4447]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Heart)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    30 µg
    Specification
    Heart
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 01 2019

    Western blot abreview for Anti-STUB1/CHIP antibody [EPR4447]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK293)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    30 µg
    Specification
    HEK293
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 01 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-STUB1/CHIP antibody [EPR4447]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (primary cortical neuron)
    Permeabilization
    Yes - 0.1% Triton/PBS 5 min
    Specification
    primary cortical neuron
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jun 23 2014

    Immunoprecipitation abreview for Anti-STUB1/CHIP antibody [EPR4447]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Mouse Cell lysate - whole cell (mouse embryonic fibroblasts)
    Total protein in input
    500 µg
    Immuno-precipitation step
    Other - Protein G Dynabeads crosslinked to antibody
    Specification
    mouse embryonic fibroblasts
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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    Verified customer

    Submitted Jun 09 2014

    Western blot abreview for Anti-STUB1/CHIP antibody [EPR4447]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (primary neurons and MEFs)
    Gel Running Conditions
    Reduced Denaturing (10% Bis-Tris)
    Loading amount
    20 µg
    Specification
    primary neurons and MEFs
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jun 09 2014

    Immunocytochemistry/ Immunofluorescence abreview for Anti-STUB1 / CHIP antibody [EPR4447]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    No
    Specification
    HeLa
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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    Verified customer

    Submitted Mar 05 2013

    Immunoprecipitation abreview for Anti-STUB1/CHIP antibody [EPR4447]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - whole cell (HeLa)
    Total protein in input
    1000 µg
    Immuno-precipitation step
    Protein A/G
    Specification
    HeLa
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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    Verified customer

    Submitted Mar 05 2013

    Western blot abreview for Anti-STUB1/CHIP antibody [EPR4447]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    HeLa
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Jan 03 2013

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