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  1. Link

    tubulin-antibody-yl12-loading-control-ab6160.pdf

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Isotype/Loading Controls Loading Controls Tubulin
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Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)

  • Datasheet
  • SDS
Reviews (37)Q&A (1)References (430)

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Immunocytochemistry/ Immunofluorescence - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)
  • Western blot - Anti-Tubulin antibody [YL1/2] (ab6160)
  • Immunocytochemistry/ Immunofluorescence - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)
  • Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)
  • Flow Cytometry - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)

Key features and details

  • Rat monoclonal [YL1/2] to Tubulin - Loading Control
  • Suitable for: WB, Flow Cyt, ICC/IF, IHC-P
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG2a

Conjugates logo Related conjugates and formulations

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Overview

  • Product name

    Anti-Tubulin antibody [YL1/2] - Loading Control
    See all Tubulin primary antibodies
  • Description

    Rat monoclonal [YL1/2] to Tubulin - Loading Control
  • Host species

    Rat
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Pig, Saccharomyces cerevisiae, Xenopus laevis, Caenorhabditis elegans, Drosophila melanogaster, Schizosaccharomyces pombe, a wide range of other species, Mammals, African green monkey
  • Immunogen

    Full length native protein (purified) corresponding to Saccharomyces cerevisiae Tubulin.

  • Epitope

    The YL1/2 monoclonal epitope has been mapped to the last 8 residues (GEEEGEEY) at the carboxy terminus of alpha tubulin when tyrosinated (PubMed IDs: 6415068, 6204858).
  • Positive control

    • ICC/IF: HeLa cells. IHC-P: Human colon tissue. WB: HeLa, NIH/3T3, BALB/3T3 and PC-12 whole cell lysate. Flow Cyt: HeLa cells.
  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • Purity

    Protein G purified
  • Primary antibody notes

    This antibody can be used as a loading control on Western blots (Allen et al.) and is not detected by anti-mouse Ig secondaries. It has been used in epitope tagging procedures to detect proteins tagged with a C-terminal Gly-Gly-Phe(OH) epitope. Under some circumstances this antibody may cross-react with other protein including E. coli rec A and oxidized actin.
  • Clonality

    Monoclonal
  • Clone number

    YL1/2
  • Isotype

    IgG2a
  • Research areas

    • Isotype/Loading Controls
    • Loading Controls
    • Tubulin
    • Tags & Cell Markers
    • Subcellular Markers
    • Cytoskeleton
    • Microtubules
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microtubules
    • Tubulin
    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Axon marker
    • Microbiology
    • Organism
    • Eukaryotic Microorganism
    • Yeast
    • Saccharomyces spp

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-Tubulin antibody [YL1/2] (ab197737)
    • HRP Anti-Tubulin antibody [YL1/2] - Loading Control (ab197740)
    • Anti-Tubulin antibody [YL1/2] - BSA and Azide free (ab264519)
  • Compatible Secondaries

    • Goat Anti-Rat IgG H&L (Alexa Fluor® 488) (ab150157)
    • Goat Anti-Rat IgG H&L (HRP) (ab205720)
  • Isotype control

    • Rat IgG2a, kappa monoclonal [RTK2758] - Isotype Control (ab18450)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab6160 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 (16)
1/5000 - 1/10000.
Flow Cyt
Use 1µg for 106 cells.

ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

ICC/IF (14)
1/1000. (see PMID: 16230461)
IHC-P (2)
Use at an assay dependent concentration.
Notes
WB
1/5000 - 1/10000.
Flow Cyt
Use 1µg for 106 cells.

ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

ICC/IF
1/1000. (see PMID: 16230461)
IHC-P
Use at an assay dependent concentration.

Target

  • Function

    Tubulin is the major constituent of microtubules. It binds two moles of GTP, one at an exchangeable site on the beta chain and one at a non-exchangeable site on the alpha-chain.
  • Sequence similarities

    Belongs to the tubulin family.
  • Post-translational
    modifications

    Undergoes a tyrosination/detyrosination cycle, the cyclic removal and re-addition of a C-terminal tyrosine residue by the enzymes tubulin tyrosine carboxypeptidase (TTCP) and tubulin tyrosine ligase (TTL), respectively.
    Some glutamate residues at the C-terminus are polyglutamylated. This modification occurs exclusively on glutamate residues and results in polyglutamate chains on the gamma-carboxyl group. Also monoglycylated but not polyglycylated due to the absence of functional TTLL10 in human. Monoglycylation is mainly limited to tubulin incorporated into axonemes (cilia and flagella) whereas glutamylation is prevalent in neuronal cells, centrioles, axonemes, and the mitotic spindle. Both modifications can coexist on the same protein on adjacent residues, and lowering glycylation levels increases polyglutamylation, and reciprocally. The precise function of such modifications is still unclear but they regulate the assembly and dynamics of axonemal microtubules.
    Acetylation of alpha-tubulins at Lys-40 stabilizes microtubules and affects affinity and processivity of microtubule motors. This modification has a role in multiple cellular functions, ranging from cell motility, cell cycle progression or cell differentiation to intracellular trafficking and signaling.
  • Cellular localization

    Cytoplasm > cytoskeleton.
  • Target information above from: UniProt accession P68363 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 10376 Human
    • Entrez Gene: 203068 Human
    • Entrez Gene: 7280 Human
    • Entrez Gene: 22143 Mouse
    • Entrez Gene: 22151 Mouse
    • Entrez Gene: 22153 Mouse
    • Entrez Gene: 500929 Rat
    • Omim: 191130 Human
    • Omim: 602530 Human
    • Omim: 615101 Human
    • SwissProt: P07437 Human
    • SwissProt: P68363 Human
    • SwissProt: Q13885 Human
    • SwissProt: P05213 Mouse
    • SwissProt: Q7TMM9 Mouse
    • SwissProt: Q9D6F9 Mouse
    • SwissProt: Q6P9V9 Rat
    • Unigene: 524390 Human
    • Unigene: 392113 Mouse
    • Unigene: 99661 Rat
    see all
  • Alternative names

    • Tubulin beta 2b antibody
    • Alpha tubulin antibody
    • Alpha-tubulin ubiquitous antibody
    • beta Ib tubulin antibody
    • CDCBM5 antibody
    • CDCBM6 antibody
    • fd02b12 antibody
    • K ALPHA 1 antibody
    • M40 antibody
    • OK/SW-cl.56 antibody
    • TBA1B_HUMAN antibody
    • TUBA1B antibody
    • TUBB1 antibody
    • TUBB2 antibody
    • TUBB5 antibody
    • tubulin alpha 1b antibody
    • Tubulin alpha-1B chain antibody
    • Tubulin alpha-ubiquitous chain antibody
    • Tubulin beta 1b antibody
    • Tubulin beta 2A antibody
    • tubulin beta 2A class IIa antibody
    • Tubulin beta antibody
    • tubulin beta chain antibody
    • tubulin beta class I antibody
    • tubulin beta-1 chain antibody
    • tubulin beta-2A chain antibody
    • tubulin beta-5 chain antibody
    • Tubulin K-alpha-1 antibody
    • tubulin, alpha, ubiquitous antibody
    • tubulin, beta 2A class IIa antibody
    • tubulin, beta polypeptide 2 antibody
    • tubulin, beta polypeptide antibody
    • tubulin, beta, class IIA antibody
    • wu:fd02b12 antibody
    • zgc:55461 antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)
    Immunocytochemistry/ Immunofluorescence - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)

    ICC/IF image of ab6160 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

    The cells were fixed in 100% methanol 5 minutes, permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6160, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was ab150165 Alexa Fluor® 488 goat anti-rat IgG (H+L) pre-adsorbed, used at a 1/1000 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    The negative control (inset) is a secondary-only assay to demonstrate low non-specific binding of the secondary antibody.

    This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10 minutes).

     

  • Western blot - Anti-Tubulin antibody [YL1/2] (ab6160)
    Western blot - Anti-Tubulin antibody [YL1/2] (ab6160)
    Lanes 1 & 3 : Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1/5000 dilution
    Lanes 2 & 4 : Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1/10000 dilution

    Lanes 1-2 : HeLa (Human epithelial carcinoma cell line) whole cell lysate
    Lanes 3-4 : BALB/3T3 whole cell lysate (ab7901)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Rabbit Anti-Rat IgG H&L (HRP) (ab6734) at 1/2000 dilution

    Predicted band size: 50 kDa
    Observed band size: 52 kDa why is the actual band size different from the predicted?
    Additional bands at: 17 kDa, 34 kDa, 80 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 10 seconds
  • Immunocytochemistry/ Immunofluorescence - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)
    Immunocytochemistry/ Immunofluorescence - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)Loison-Robert et al PLoS One. 2018 Jan 25;13(1):e0190014. doi: 10.1371/journal.pone.0190014. eCollection 2018. Fig 5. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Cytoskeleton and major extracellular matrix proteins in human DPSC (Dental pulp stem cell) were analyzed by immunofluorescence.

    Vimentin and tubulin (Panel D, control, E, (BD) and F, (BR)) are shown.

    After 7 days in contact with/without the materials (Biodentine (BD) and Bioroot (BR)), coated coverslip cultures were fixed in PBS (pH 7.4) containing 4% paraformaldehyde/5% sucrose for 10 minutes. For detection of intracellular molecules, the cells on the coverslips were permeabilized using 0.5% Triton X-100. To block background staining, cells were treated with PBS containing 1% BSA/1% glycine at 37°C for 20 minutes. Samples were incubated with the primary antibody at 4°C overnight or at 37°C for 2 hours. For double immunostaining, primary antibodies were incubated as above. Samples were then incubated with the appropriate secondary antibodies at 37°C for 1 hour. Cell nuclei were stained using DAPI.

    Scale Bar: 100 μm.

  • Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)
    Western blot - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)
    All lanes : Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) whole cell lysate
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
    Lane 3 : PC-12 (Rat adrenal pheochromocytoma cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Peroxidase Conjugated Rabbit Anti-Rat IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 50 kDa
    Observed band size: 52 kDa why is the actual band size different from the predicted?
    Additional bands at: 85 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 8 minutes
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)

    IHC image of Tubulin staining in human colon formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer pH 6 for 20 minutes. The section was then incubated with ab6160, 5 µg/ml, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Flow Cytometry - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)
    Flow Cytometry - Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160)

    Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with ab6160 (red line).

    The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab6160, 1 µg/1x106 cells) for 30 minutes at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was rat IgG2a [aRTK2758] (ab18450, 1 µg/1x106 cells) used under the same conditions.

    Acquisition of >5,000 events was performed.

Protocols

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

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (430)

Publishing research using ab6160? Please let us know so that we can cite the reference in this datasheet.

ab6160 has been referenced in 430 publications.

  • Gori I  et al. Mutations in SKI in Shprintzen-Goldberg syndrome lead to attenuated TGF-ß responses through SKI stabilization. Elife 10:N/A (2021). PubMed: 33416497
  • McGinley LM  et al. Magnetic resonance imaging of human neural stem cells in rodent and primate brain. Stem Cells Transl Med 10:83-97 (2021). PubMed: 32841522
  • Su F  et al. Recombinant high-mobility group box 1 induces cardiomyocyte hypertrophy by regulating the 14-3-3?, PI3K and nuclear factor of activated T cells signaling pathways. Mol Med Rep 23:N/A (2021). PubMed: 33495819
  • Pesaola F  et al. The neuronal ceroid lipofuscinosis-related protein CLN8 regulates endo-lysosomal dynamics and dendritic morphology. Biol Cell N/A:N/A (2021). PubMed: 34021618
  • Zhang H  et al. Hepcidin-induced reduction in iron content and PGC-1ß expression negatively regulates osteoclast differentiation to play a protective role in postmenopausal osteoporosis. Aging (Albany NY) 13:11296-11314 (2021). PubMed: 33820875
View all Publications for this product

Customer reviews and Q&As

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1-10 of 37 Abreviews

Western blot abreview for Anti-Tubulin antibody [YL1/2] - Loading Control

Excellent
Abreviews
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (lung, pancreatic, placenta)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
lung, pancreatic, placenta
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°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 Sep 07 2021

Immunoprecipitation abreview for Anti-Tubulin antibody [YL1/2] - Loading Control

Excellent
Abreviews
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (293T cells)
Total protein in input
500 µg
Immuno-precipitation step
Protein A/G
Specification
293T cells
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 Nov 08 2019

Western blot abreview for Anti-Tubulin antibody [YL1/2]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (293T cells)
Gel Running Conditions
Reduced Denaturing (4-12% Gradient Gel)
Loading amount
50 µg
Specification
293T cells
Blocking step
Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · 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 Sep 10 2018

Immunoprecipitation abreview for Anti-Tubulin antibody [YL1/2]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (MEF cells)
Total protein in input
500 µg
Immuno-precipitation step
Protein A/G
Specification
MEF cells
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 Sep 06 2018

Immunocytochemistry/ Immunofluorescence abreview for Anti-Tubulin antibody [YL1/2]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (MEFs)
Permeabilization
Yes - Digitonin
Specification
MEFs
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 27°C
Fixative
Formaldehyde
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 31 2018

Western blot abreview for Anti-Tubulin antibody [YL1/2]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEF cells)
Gel Running Conditions
Reduced Denaturing (4-12% Gradient Gel)
Loading amount
50 µg
Specification
MEF cells
Blocking step
Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · 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 Aug 01 2018

Western blot abreview for Anti-Tubulin antibody [YL1/2]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (brain cortex)
Gel Running Conditions
Reduced Denaturing (8%)
Loading amount
30 µg
Specification
brain cortex
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 22°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.

DR. Alexandre Magno

Verified customer

Submitted Jul 23 2018

Western blot abreview for Anti-Tubulin antibody [YL1/2]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Human cerebral organoid)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (12.5% gel)
Loading amount
20 µg
Specification
Human cerebral organoid
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 May 30 2018

Immunocytochemistry/ Immunofluorescence abreview for Anti-Tubulin antibody [YL1/2]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Trophoblast giant cells)
Permeabilization
No
Specification
Trophoblast giant cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: 20°C
Fixative
Methanol
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.

MISS. Stephanie Chrysanthou

Verified customer

Submitted Jul 10 2017

Western blot abreview for Anti-Tubulin antibody [YL1/2]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - other (Hippocampus)
Gel Running Conditions
Reduced Denaturing (13)
Loading amount
6 µg
Specification
Hippocampus
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.

DR. Sergi Bayod

Verified customer

Submitted Nov 28 2016

1-10 of 37 Abreviews

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