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  1. Link

    atg7-antibody-epr6251-ab133528.pdf

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Cell Biology Apoptosis Intracellular Associated Proteins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ATG7 antibody [EPR6251] (ab133528)

  • Datasheet
  • SDS
Reviews (16) Submit a question References (49)

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Western blot - Anti-ATG7 antibody [EPR6251] (ab133528)
  • Immunocytochemistry/ Immunofluorescence - Anti-ATG7 antibody [EPR6251] (ab133528)
  • Western blot - Anti-ATG7 antibody [EPR6251] (ab133528)
  • Western blot - Anti-ATG7 antibody [EPR6251] (ab133528)
  • Western blot - Anti-ATG7 antibody [EPR6251] (ab133528)
  • Immunocytochemistry/ Immunofluorescence - Anti-ATG7 antibody [EPR6251] (ab133528)
  • Anti-ATG7 antibody [EPR6251] (ab133528)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6251] to ATG7
  • Suitable for: WB, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Carrier Free HRP

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Overview

  • Product name

    Anti-ATG7 antibody [EPR6251]
    See all ATG7 primary antibodies
  • Description

    Rabbit monoclonal [EPR6251] to ATG7
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human ATG7 aa 1-100. The exact sequence is proprietary.

  • Positive control

    • 293T, HepG2 and Jurkat whole cell lysate (ab7899), Rat spleen and kidney tissue lysates, mouse spleen and kidney tissue lysates; HT-29 and HeLa cells.
  • General notes

    We have had 1 attempt at IHC-P with ab133528 in our own lab. We observed both cytoplasmic and nuclear staining on several tissues (including human stomach, kidney and pancreatic cancer), under our experimental conditions. For IHC-P on human tissues, we would recommend using ab52472.

    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. Avoid freeze / thaw cycle. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR6251
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Intracellular
    • Associated Proteins
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ub-like Proteins
    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Cardiovascular
    • Heart
    • Autophagy
    • APG gene products
    • Cancer
    • Signal transduction
    • Autophagy
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitophagy fission and fusion
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Autophagy and mitophagy
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Autophagy and mitophagy
    • APG gene products
    • Cancer
    • Cell Death
    • Autophagy
    • Signal Transduction
    • Cancer
    • Cell Death
    • Autophagy
    • APG gene products
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-ATG7 antibody [EPR6251] (ab214867)
    • HRP Anti-ATG7 antibody [EPR6251] (ab215336)
    • Anti-ATG7 antibody [EPR6251] - BSA and Azide free (ab232348)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

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

    • Jurkat cytoplasmic extract lysate (ab14842)
    • Mouse kidney normal tissue lysate - total protein (ab29305)
    • Mouse spleen tissue lysate - total protein (ab7937)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133528 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 (9)
1/10000 - 1/50000. Predicted molecular weight: 77 kDa.

Use 5% non-fat dry milk + TBST for blocking.

ICC/IF (1)
1/100 - 1/500.
Notes
WB
1/10000 - 1/50000. Predicted molecular weight: 77 kDa.

Use 5% non-fat dry milk + TBST for blocking.

ICC/IF
1/100 - 1/500.

Target

  • Function

    E1-like activating enzyme involved in the 2 ubiquitin-like systems required for cytoplasm to vacuole transport (Cvt) and autophagy. Activates ATG12 for its conjugation with ATG5 as well as the ATG8 family proteins for their conjugation with phosphatidylethanolamine. Both systems are needed for the ATG8 association to Cvt vesicles and autophagosomes membranes. Required for autophagic death induced by caspase-8 inhibition. Required for mitophagy which contributes to regulate mitochondrial quantity and quality by eliminating the mitochondria to a basal level to fulfill cellular energy requirements and preventing excess ROS production. Modulates p53/TP53 activity to regulate cell cycle and survival during metabolic stress. Plays also a key role in the maintenance of axonal homeostasis, the prevention of axonal degeneration, the maintenance of hematopoietic stem cells, the formation of Paneth cell granules, as well as in adipose differentiation.
  • Tissue specificity

    Widely expressed, especially in kidney, liver, lymph nodes and bone marrow.
  • Sequence similarities

    Belongs to the ATG7 family.
  • Domain

    The C-terminal part of the protein is essential for the dimerization and interaction with ATG3 and ATG12.
    The N-terminal FAP motif (residues 15 to 17) is essential for the formation of the ATG89-PE and ATG5-ATG12 conjugates.
  • Post-translational
    modifications

    Acetylated by EP300.
  • Cellular localization

    Cytoplasm. Preautophagosomal structure. Localizes also to discrete punctae along the ciliary axoneme and to the base of the ciliary axoneme.
  • Target information above from: UniProt accession O95352 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 10533 Human
    • Entrez Gene: 74244 Mouse
    • Entrez Gene: 312647 Rat
    • Omim: 608760 Human
    • SwissProt: O95352 Human
    • SwissProt: Q9D906 Mouse
    • SwissProt: Q641Y5 Rat
    • Unigene: 38032 Human
    • Unigene: 275332 Mouse
    • Unigene: 162765 Rat
    see all
  • Alternative names

    • 1810013K23Rik antibody
    • Apg 7 antibody
    • APG7 autophagy 7 like antibody
    • APG7 autophagy 7-like (S. cerevisiae) antibody
    • APG7 like antibody
    • APG7, S. cerevisiae, homolog of antibody
    • APG7-like antibody
    • APG7L antibody
    • ATG 7 antibody
    • ATG12-activating enzyme E1 ATG7 antibody
    • ATG7 antibody
    • ATG7 autophagy related 7 homolog (S. cerevisiae) antibody
    • ATG7 autophagy related 7 homolog antibody
    • ATG7_HUMAN antibody
    • Atg7l antibody
    • Autophagy 7, S. cerevisiae, homolog of antibody
    • Autophagy related protein 7 antibody
    • Autophagy-related 7 (yeast) antibody
    • Autophagy-related protein 7 antibody
    • DKFZp434N0735 antibody
    • GSA 7 antibody
    • GSA7 antibody
    • hAGP7 antibody
    • Ubiquitin activating enzyme E1 like protein antibody
    • Ubiquitin-activating enzyme E1-like protein antibody
    • Ubiquitin-like modifier-activating enzyme ATG7 antibody
    see all

Images

  • Western blot - Anti-ATG7 antibody [EPR6251] (ab133528)
    Western blot - Anti-ATG7 antibody [EPR6251] (ab133528)
    All lanes : Anti-ATG7 antibody [EPR6251] (ab133528) at 1/10000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ATG7 knockout HeLa cell lysate
    Lane 3 : HepG2 cell lysate
    Lane 4 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 77 kDa
    Observed band size: 75 kDa why is the actual band size different from the predicted?



    False colour image of Western blot: Anti-ATG7 antibody [EPR6251] staining at 1/10000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab133528 was shown to bind specifically to ATG7. A band was observed at 75 kDa in wild-type HeLa cell lysates with no signal observed at this size in ATG7 knockout cell line ab283307 (knockout cell lysate ab287353). To generate this image, wild-type and ATG7 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-ATG7 antibody [EPR6251] (ab133528)
    Immunocytochemistry/ Immunofluorescence - Anti-ATG7 antibody [EPR6251] (ab133528)

    Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ATG7 with purified ab133528 at 1/150 dilution (8.5μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889  Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1/1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Western blot - Anti-ATG7 antibody [EPR6251] (ab133528)
    Western blot - Anti-ATG7 antibody [EPR6251] (ab133528)
    All lanes : Anti-ATG7 antibody [EPR6251] (ab133528) at 1/50000 dilution (purified)

    Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 2 : Mouse spleen lysate
    Lane 3 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 4 : Mouse kidney lysate
    Lane 5 : Rat kidney lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 77 kDa
    Observed band size: 77 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Western blot - Anti-ATG7 antibody [EPR6251] (ab133528)
    Western blot - Anti-ATG7 antibody [EPR6251] (ab133528)
    All lanes : Anti-ATG7 antibody [EPR6251] (ab133528) at 1/10000 dilution (purified)

    Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 2 : Rat spleen lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 77 kDa
    Observed band size: 77 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Western blot - Anti-ATG7 antibody [EPR6251] (ab133528)
    Western blot - Anti-ATG7 antibody [EPR6251] (ab133528)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: ATG7 knockout HAP1 cell lysate (20 µg)
    Lane 3: Jurkat cell lysate (20 µg)
    Lane 4: HepG2 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab133528 observed at 77 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab133528 was shown to specifically react with ATG7 when ATG7 knockout samples were used. Wild-type and Apg7 knockout samples were subjected to SDS-PAGE. ab133528 and ab8245 (loading control to GAPDH) were diluted 1/10,000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-ATG7 antibody [EPR6251] (ab133528)
    Immunocytochemistry/ Immunofluorescence - Anti-ATG7 antibody [EPR6251] (ab133528)

    Immunocytochemistry/Immunofluorescence analysis of HT-29 (Human colorectal adenocarcinoma cell line) labeling ATG7 with purified ab133528 at 1/500 dilution. Cells were fixed with 100% methanol. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

  • Anti-ATG7 antibody [EPR6251] (ab133528)
    Anti-ATG7 antibody [EPR6251] (ab133528)

Protocols

  • Western blot protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (49)

Publishing research using ab133528? Please let us know so that we can cite the reference in this datasheet.

ab133528 has been referenced in 49 publications.

  • Wang J  et al. Activation of CREB-mediated autophagy by thioperamide ameliorates ß-amyloid pathology and cognition in Alzheimer's disease. Aging Cell 20:e13333 (2021). PubMed: 33682314
  • Zhang Z  et al. CircRAB11FIP1 promoted autophagy flux of ovarian cancer through DSC1 and miR-129. Cell Death Dis 12:219 (2021). PubMed: 33637694
  • Luu AZ  et al. Loss of endothelial cell-specific autophagy-related protein 7 exacerbates doxorubicin-induced cardiotoxicity. Biochem Biophys Rep 25:100926 (2021). PubMed: 33553688
  • Zhang J  et al. TGF-ß1-induced autophagy activates hepatic stellate cells via the ERK and JNK signaling pathways. Int J Mol Med 47:256-266 (2021). PubMed: 33236148
  • Chen J  et al. Inhibition of Autophagy Prevents Panax Notoginseng Saponins (PNS) Protection on Cardiac Myocytes Against Endoplasmic Reticulum (ER) Stress-Induced Mitochondrial Injury, Ca2+ Homeostasis and Associated Apoptosis. Front Pharmacol 12:620812 (2021). PubMed: 33762943
View all Publications for this product

Customer reviews and Q&As

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1-10 of 16 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-ATG7 antibody [EPR6251]

Inconclusive
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Common marmoset Tissue sections (Pancreas)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Pancreas
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°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.

MR. Carl Hobbs

Verified customer

Submitted Aug 17 2018

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-ATG7 antibody [EPR6251]

Inconclusive
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°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.

MR. Carl Hobbs

Verified customer

Submitted Aug 17 2018

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-ATG7 antibody [EPR6251]

Inconclusive
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Liver
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°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.

MR. Carl Hobbs

Verified customer

Submitted Aug 17 2018

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-ATG7 antibody [EPR6251]

Inconclusive
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Brain
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°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.

MR. Carl Hobbs

Verified customer

Submitted Aug 17 2018

Western blot abreview for Anti-Apg7 antibody [EPR6251]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 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 Feb 08 2018

Western blot abreview for Anti-Apg7 antibody [EPR6251]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Rat Cell lysate - whole cell (Cardiomyocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
Cardiomyocytes
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 Feb 08 2018

Western blot abreview for Anti-Apg7 antibody [EPR6251]

Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Cardiomyocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
Cardiomyocytes
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 Feb 01 2018

Immunocytochemistry abreview for Anti-Apg7 antibody [EPR6251]

Average
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Cardiomyocytes)
Permeabilization
Yes - Triton x-100, 0.03%
Specification
Cardiomyocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°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 Jan 31 2018

Western blot abreview for Anti-Apg7 antibody [EPR6251]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Rhesus monkey Tissue lysate - whole (Heart)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 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 Jan 30 2018

Western blot abreview for Anti-Apg7 antibody [EPR6251]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Kidney)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
Kidney
Blocking step
BSA 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 19 2017

1-10 of 16 Abreviews or Q&A

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