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    gapdh-antibody-epr16884-loading-control-ab181603.pdf

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RecombinantRabMAb

Recombinant Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (6) Submit a question References (146)

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Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
  • Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
  • Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
  • Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
  • Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
  • Flow Cytometry (Intracellular) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
  • Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR16884] to GAPDH - Loading Control
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Reacts with: Mouse, Rat, Chicken, Cow, Dog, Human, African green monkey

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free HRP PE

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Overview

  • Product name

    Anti-GAPDH antibody [EPR16884] - Loading Control
    See all GAPDH primary antibodies
  • Description

    Rabbit monoclonal [EPR16884] to GAPDH - Loading Control
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Chicken, Cow, Dog, Human, African green monkey
    Predicted to work with: Monkey
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, MDBK, COS-1, MDCK, UMNSAH/DF-1, Jurkat, C6 and NIH/3T3 whole cell lysates; Mouse brain and heart lysates; Rat brain, heart, kidney and spleen lysates; Human fetal brain, heart and kidney lysates. IHC-P: Human transitional cell carcinoma of bladder, Mouse spleen and Rat spleen tissues. ICC/IF: HeLa cells. Flow: Jurkat cells.
  • 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

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

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR16884
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Huntington's disease
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Other
    • Isotype/Loading Controls
    • Loading Controls
    • GAPDH
    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of carbohydrates
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Carbohydrate metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
    • Metabolism
    • Types of disease
    • Neurodegenerative disease

Associated products

  • Alternative Versions

    • Anti-GAPDH antibody [EPR16884] - BSA and Azide free (ab199554)
    • Alexa Fluor® 488 Anti-GAPDH antibody [EPR16884] (ab204276)
    • Alexa Fluor® 647 Anti-GAPDH antibody [EPR16884] (ab204480)
    • HRP Anti-GAPDH antibody [EPR16884] - Loading Control (ab204481)
    • Alexa Fluor® 594 Anti-GAPDH antibody [EPR16884] (ab206373)
    • Alexa Fluor® 555 Anti-GAPDH antibody [EPR16884] (ab206629)
    • PE Anti-GAPDH antibody [EPR16884] (ab209037)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • 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)
  • Recombinant Protein

    • Recombinant human GAPDH protein (Active) (ab82633)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab181603 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
Flow Cyt (Intra)
1/200.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB (3)
1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF (3)
1/250.
Notes
Flow Cyt (Intra)
1/200.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/250.

Target

  • Function

    Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC (By similarity). Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate.
  • Pathway

    Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
  • Sequence similarities

    Belongs to the glyceraldehyde-3-phosphate dehydrogenase family.
  • Post-translational
    modifications

    S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
    ISGylated.
  • Cellular localization

    Cytoplasm > cytosol. Nucleus. Cytoplasm > perinuclear region. Membrane. Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions.
  • Target information above from: UniProt accession P04406 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 374193 Chicken
    • Entrez Gene: 281181 Cow
    • Entrez Gene: 403755 Dog
    • Entrez Gene: 2597 Human
    • Entrez Gene: 100042025 Mouse
    • Entrez Gene: 14433 Mouse
    • Entrez Gene: 24383 Rat
    • Entrez Gene: 685186 Rat
    • Omim: 138400 Human
    • SwissProt: P00356 Chicken
    • SwissProt: P10096 Cow
    • SwissProt: Q28259 Dog
    • SwissProt: P04406 Human
    • SwissProt: P16858 Mouse
    • SwissProt: P04797 Rat
    • Unigene: 544577 Human
    • Unigene: 592355 Human
    • Unigene: 598320 Human
    • Unigene: 304088 Mouse
    • Unigene: 309092 Mouse
    • Unigene: 317779 Mouse
    • Unigene: 343110 Mouse
    • Unigene: 392463 Mouse
    • Unigene: 392480 Mouse
    • Unigene: 414470 Mouse
    • Unigene: 458138 Mouse
    • Unigene: 458416 Mouse
    • Unigene: 475698 Mouse
    • Unigene: 129558 Rat
    • Unigene: 91450 Rat
    see all
  • Alternative names

    • 38 kDa BFA-dependent ADP-ribosylation substrate antibody
    • aging associated gene 9 protein antibody
    • Aging-associated gene 9 protein antibody
    • BARS-38 antibody
    • cb609 antibody
    • EC 1.2.1.12 antibody
    • Epididymis secretory sperm binding protein Li 162eP antibody
    • G3P_HUMAN antibody
    • G3PD antibody
    • G3PDH antibody
    • GAPD antibody
    • GAPDH antibody
    • Glyceraldehyde 3 phosphate dehydrogenase antibody
    • glyceraldehyde 3-PDH antibody
    • Glyceraldehyde-3-phosphate dehydrogenase antibody
    • HEL-S-162eP antibody
    • KNC-NDS6 antibody
    • MGC102544 antibody
    • MGC102546 antibody
    • MGC103190 antibody
    • MGC103191 antibody
    • MGC105239 antibody
    • MGC127711 antibody
    • MGC88685 antibody
    • OCAS, p38 component antibody
    • OCT1 coactivator in S phase, 38-KD component antibody
    • peptidyl cysteine S nitrosylase GAPDH antibody
    • Peptidyl-cysteine S-nitrosylase GAPDH antibody
    • wu:fb33a10 antibody
    see all

Images

  • Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
    Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
    All lanes : Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603) at 1/10000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
    Lane 2 : MDBK (Bovine kidney cell line) whole cell lysates
    Lane 3 : COS-1 (African green monkey kidney fibroblast-like cell line) whole cell lysates
    Lane 4 : MDCK (Canine kidney cell line) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 36 kDa
    Observed band size: 36 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
    Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
    All lanes : Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603) at 1/50000 dilution

    Lane 1 : UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) whole cell lysates
    Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 36 kDa
    Observed band size: 36 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
    Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
    All lanes : Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603) at 1/10000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Rat brain lysate
    Lane 4 : Rat heart lysate
    Lane 5 : Rat kidney lysate
    Lane 6 : Rat spleen lysate
    Lane 7 : C6 (Rat glial tumor cells) whole cell lysates
    Lane 8 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 36 kDa
    Observed band size: 36 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
    Western blot - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
    All lanes : Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603) at 1/10000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 36 kDa
    Observed band size: 36 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
    Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling GAPDH with ab181603 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm and nuclear staining on HeLa cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows;
    1. ab181603 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)

    Immunohistochemical analysis of paraffin-embedded Human transitional cell carcinoma of bladder tissue labeling GAPDH with ab181603 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic and nucleus staining on the tumor cells of transitional cell carcinoma of Human bladder is observed. Counter stained with Hematoxylin.

     

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)

    Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling GAPDH with ab181603 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocytes of mouse spleen is observed. Counter stained with Hematoxylin.

     

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)

    Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling GAPDH with ab181603 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocyte of rat spleen is observed. Counter stained with Hematoxylin.

     

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Flow Cytometry (Intracellular) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
    Flow Cytometry (Intracellular) - Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)

    Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling GAPDH with ab181603 at 1/200 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody. 

     

     

  • Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)
    Anti-GAPDH antibody [EPR16884] - Loading Control (ab181603)

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

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (146)

Publishing research using ab181603? Please let us know so that we can cite the reference in this datasheet.

ab181603 has been referenced in 146 publications.

  • Tian L  et al. Long non-coding RNA LINC01006 exhibits oncogenic properties in cervical cancer by functioning as a molecular sponge for microRNA-28-5p and increasing PAK2 expression. Int J Mol Med 47:N/A (2021). PubMed: 33576457
  • Azizidoost SH  et al. Impact of Methyl-ß-Cyclodextrin and Apolipoprotein A-I on The Expression of ATP-Binding Cassette Transporter A1 and Cholesterol Depletion in C57BL/6 Mice Astrocytes. Cell J 23:93-98 (2021). PubMed: 33650825
  • Yu H  et al. LINC00504 Promotes the Malignant Biological Behavior of Breast Cancer Cells by Upregulating HMGB3 via Decoying MicroRNA-876-3p. Cancer Manag Res 13:1803-1815 (2021). PubMed: 33654429
  • Tian T  et al. FAST1 Predicts Poor Survival of Renal Carcinoma and Promotes Its Progression Through the TGF-ß/Smad Pathway. Onco Targets Ther 14:1487-1499 (2021). PubMed: 33679133
  • Mosqueira M  et al. Cardiomyocyte depolarization triggers NOS-dependent NO transient after calcium release, reducing the subsequent calcium transient. Basic Res Cardiol 116:18 (2021). PubMed: 33728868
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
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1-6 of 6 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-GAPDH antibody [EPR16884]

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (undifferentiated Human ES cells on MEFs)
Permeabilization
Yes - see blocking buffer
Specification
undifferentiated Human ES cells on MEFs
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Apr 14 2016

Western blot abreview for Anti-GAPDH antibody [EPR16884]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HUES7 ES cells)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Specification
HUES7 ES cells
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
Read More

Abcam user community

Verified customer

Submitted Feb 04 2016

Immunocytochemistry/ Immunofluorescence abreview for Anti-GAPDH antibody [EPR16884]

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HEPG2)
Specification
HEPG2
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Nov 18 2015

Western blot abreview for Anti-GAPDH antibody [EPR16884]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEPG2)
Gel Running Conditions
Reduced Denaturing (12.5)
Loading amount
20 µg
Specification
HEPG2
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
Read More

Abcam user community

Verified customer

Submitted Nov 13 2015

Immunocytochemistry/ Immunofluorescence abreview for Anti-GAPDH antibody [EPR16884]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 16°C
Sample
Mouse Cell (mouse embryonic stem cell)
Specification
mouse embryonic stem cell
Permeabilization
Yes - 0.1% triton X
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Aug 18 2014

Western blot abreview for Anti-GAPDH antibody [EPR16884]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Loading amount
100000 cells
Gel Running Conditions
Reduced Denaturing (10% gel)
Sample
Mouse Cell lysate - whole cell (mouse embryonic stem cell)
Specification
mouse embryonic stem cell
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
Read More

Abcam user community

Verified customer

Submitted Aug 08 2014

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