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  1. Link

    products/primary-antibodies/cox-iv-antibody-mabcam33985-mitochondrial-marker-ab33985.pdf

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Tags & Cell Markers Subcellular Markers Organelles Mitochondria
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Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)

  • Datasheet
  • SDS
Reviews (25)Q&A (1)References (90)

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Western blot - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)
  • Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)
  • Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)
  • Flow Cytometry - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)
  • Western blot - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)
  • Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)
  • Western blot - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)

Key features and details

  • Mouse monoclonal [mAbcam33985] to COX IV - Mitochondrial Marker
  • Suitable for: Flow Cyt, WB, ICC/IF
  • Reacts with: Mouse, Cow, Human, Xenopus laevis
  • Isotype: IgG1

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 HRP

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Protein
Product image
Recombinant Human COX IV protein (ab112273)
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FITC Anti-DNA/RNA Damage antibody [15A3] (ab183393)
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Anti-TOMM20 antibody [EPR15581-39] - Mitochondrial Marker (ab186734)

View more associated products

Overview

  • Product name

    Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker
    See all COX IV primary antibodies
  • Description

    Mouse monoclonal [mAbcam33985] to COX IV - Mitochondrial Marker
  • Host species

    Mouse
  • Tested applications

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

    Reacts with: Mouse, Cow, Human, Xenopus laevis
    Predicted to work with: Rat, Sheep, Chimpanzee, Monkey, Zebrafish, African green monkey, Chinese hamster, Drosophila C virus
  • Immunogen

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

  • Positive control

    • WB: Jurkat and HepG2 whole cell lysates and human skeletal muscle, mouse skeletal muscle and cow kidney tissue lysates. ICC/IF: HeLa cells; 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
  • Clonality

    Monoclonal
  • Clone number

    mAbcam33985
  • Myeloma

    Sp2
  • Isotype

    IgG1
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Organelles
    • Mitochondria
    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Isotype/Loading Controls
    • Loading Controls
    • COX IV - Mitochondrial
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Integration of energy metabolism
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Integration of energy
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Oxidative phosphorylation
    • Complex IV
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Alternative Versions

    • Alexa Fluor® 647 Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab197491)
    • HRP Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab62164)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Recombinant Protein

    • Recombinant Human COX IV protein (ab112273)
  • Related Products

    • Mitochondrial Staining Kit - Red Fluorescence - Cytopainter (ab112145)
    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)
    • Mitochondrial Staining Kit - NIR Fluorescence - Cytopainter (ab176747)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab33985 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
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB (13)
Use a concentration of 1 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
ICC/IF (9)
Use a concentration of 1 µg/ml.
Notes
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB
Use a concentration of 1 µg/ml. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
ICC/IF
Use a concentration of 1 µg/ml.

Target

  • Function

    This protein is one of the nuclear-coded polypeptide chains of cytochrome c oxidase, the terminal oxidase in mitochondrial electron transport.
  • Tissue specificity

    Ubiquitous.
  • Sequence similarities

    Belongs to the cytochrome c oxidase IV family.
  • Cellular localization

    Mitochondrion inner membrane.
  • Target information above from: UniProt accession P13073 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 454391 Chimpanzee
    • Entrez Gene: 1327 Human
    • Entrez Gene: 12857 Mouse
    • Entrez Gene: 29445 Rat
    • Entrez Gene: 326975 Zebrafish
    • Omim: 123864 Human
    • SwissProt: O46577 Chimpanzee
    • SwissProt: P00423 Cow
    • SwissProt: P13073 Human
    • SwissProt: P19783 Mouse
    • SwissProt: P10888 Rat
    • Unigene: 433419 Human
    • Unigene: 386758 Mouse
    • Unigene: 2528 Rat
    see all
  • Alternative names

    • AL024441 antibody
    • COX 4 antibody
    • COX IV 1 antibody
    • COX IV antibody
    • COX IV-1 antibody
    • Cox4 antibody
    • COX41_HUMAN antibody
    • Cox4a antibody
    • COX4B antibody
    • COX4I1 antibody
    • COX4I2 antibody
    • COX4L2 antibody
    • COXIV antibody
    • Cytochrome c oxidase polypeptide IV antibody
    • Cytochrome c oxidase subunit 4 isoform 1 mitochondrial antibody
    • Cytochrome c oxidase subunit 4 isoform 1, mitochondrial antibody
    • Cytochrome C Oxidase subunit IV antibody
    • Cytochrome c oxidase subunit IV isoform 1 antibody
    • Cytochrome c oxidase subunit IV isoform 2 (lung) antibody
    • Cytochrome c oxydase subunit 4 antibody
    • dJ857M17.2 antibody
    • MGC105470 antibody
    • MGC72016 antibody
    see all

Images

  • Western blot - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)
    Western blot - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)
    All lanes : Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985) at 1 µg/ml

    Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 3 : Human skeletal muscle tissue lysate - total protein (ab29330)
    Lane 4 : Skeletal Muscle (Mouse) Tissue Lysate
    Lane 5 : Kidney (Cow) Tissue Lysate (ab29073)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 15 kDa
    Observed band size: 15 kDa


    Exposure time: 1 minute
  • Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)
    Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)

    Ab33985 staining COX IV in HeLa (Human cervix adenocarcinoma epithelial cell) cells by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% TritonX-100. Samples were incubated with primary antibody at 1:1000 dilution (1µg/ml). An Alexa Fluor®594 Goat anti-mouse (ab150120) was used as a secondary antibody at 1:1000 dilution (2 µg/ml). Cells were counterstained with anti-Cyclophilin F (ab231155, 5.5µg/ml) and AlexaFluor®488 Goat anti-Rabbit (ab150077, 2µg/ml).  DAPI was used as a nuclear counterstain. Ab33985 was used for negative control 1 at 1:1000 dilution (1µg/ml). For negative control 2, ab231155 was used at a 1:100 dilution (5.5µg/ml) and ab150129 was used as a secondary antibody at 1:1000 dilution (2µg/ml). Confocal image showing mitochondrial staining in HeLa cell line. 

  • Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)
    Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)

    ICC/IF image of ab33985 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab33985, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

  • Flow Cytometry - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)
    Flow Cytometry - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)

    Overlay histogram showing HeLa cells stained with ab33985 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. 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 (ab33985, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Western blot - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)
    Western blot - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)This image is courtesy of an Abreview submitted by Dr Anne-Lore Schlaitz
    All lanes : Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985) at 1/1000 dilution

    Lane 1 : Crude extract prepared from Xenopus laevis egg
    Lane 2 : Cytosol lysate prepared from Xenopus laevis egg extract
    Lane 3 : Total membrane lysate prepared from Xenopus laevis egg extract

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : HRP conjugated donkey anti-mouse IgG at 1/4000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 15 kDa
    Observed band size: 15 kDa


    Exposure time: 90 minutes

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)
    Immunocytochemistry/ Immunofluorescence - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)This image is courtesy of an anonymous Abreview

    ab33985 staining COX IV in human proximal tubular epithelial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 in PBS and blocked with 3% BSA for 15 minutes at 20°C. Samples were incubated with primary antibody (1/200 in PBS) for 45 minutes at 20°C. ab6785, a FITC-conjugated goat anti-mouse IgG (H+L) polyclonal was used as the secondary antibody (1/1000).

    See Abreview

  • Western blot - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)
    Western blot - Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985)This image is courtesy of an anonymous Abreview
    All lanes : Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985) at 1/1000 dilution

    All lanes : cow liver membrane with Milk

    Lysates/proteins at 15 µg per lane.

    Blocking peptides at 1.5 % per lane.

    Secondary
    All lanes : Goat Anti-mouse IgG HRP at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 15 kDa


    Exposure time: 17 hours

    See Abreview

Protocols

  • Mouse on Mouse staining protocol

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (90)

Publishing research using ab33985? Please let us know so that we can cite the reference in this datasheet.

ab33985 has been referenced in 90 publications.

  • Filippou A  et al. ANO1 Expression Orchestrates p27Kip1/MCL1-Mediated Signaling in Head and Neck Squamous Cell Carcinoma. Cancers (Basel) 13:N/A (2021). PubMed: 33803266
  • Meng L  et al. Dysregulation of the Sirt5/IDH2 axis contributes to sunitinib resistance in human renal cancer cells. FEBS Open Bio 11:921-931 (2021). PubMed: 33455080
  • Choi S  et al. Suppression of Foxo3-Gatm by miR-132-3p Accelerates Cyst Formation by Up-Regulating ROS in Autosomal Dominant Polycystic Kidney Disease. Biomol Ther (Seoul) 29:311-320 (2021). PubMed: 33408288
  • Cheng Y  et al. iTRAQ-Based Quantitative Proteomics Indicated Nrf2/OPTN-Mediated Mitophagy Inhibits NLRP3 Inflammasome Activation after Intracerebral Hemorrhage. Oxid Med Cell Longev 2021:6630281 (2021). PubMed: 33628368
  • Duong A  et al. Characterization of mitochondrial health from human peripheral blood mononuclear cells to cerebral organoids derived from induced pluripotent stem cells. Sci Rep 11:4523 (2021). PubMed: 33633238
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-10 of 26 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Zebrafish Cell (Brain)
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 26°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.

DR. Haibo Li

Verified customer

Submitted Feb 26 2020

Western blot abreview for Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Hematological cancer cell line lysates)
Gel Running Conditions
Reduced Denaturing (15)
Loading amount
20 µg
Specification
Hematological cancer cell line lysates
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · 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 14 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Zebrafish Cell (Mesenchyme)
Permeabilization
Yes - 0.3% Triton X-100
Specification
Mesenchyme
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°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 Dec 27 2018

Western blot abreview for Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Cow Tissue lysate - other (Liver membrane)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
30 µg
Specification
Liver membrane
Blocking step
Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 1.5% · 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 05 2018

Immunocytochemistry/ Immunofluorescence abreview for Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.1% Triton-X100 in PBS
Specification
HeLa
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · 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 Jul 28 2017

Western blot abreview for Anti-COX IV [mAbcam33985] antibody

Excellent
Abreviews
Abreviews
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (10% SDS)
Sample
Mouse Tissue lysate - whole (heart)
Specification
heart
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°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 16 2013

Western blot abreview for Anti-COX IV [mAbcam33985] antibody

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Sheep Tissue lysate - whole (liver)
Loading amount
30 µg
Specification
liver
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 1.5% · 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 Jul 12 2011

Immunocytochemistry/ Immunofluorescence abreview for Anti-COX IV [mAbcam33985] antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (leukocytes from bone marrow)
Specification
leukocytes from bone marrow
Fixative
Methanol
Permeabilization
No
Blocking step
Serum as blocking agent for 2 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 27 2011

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-COX IV [mAbcam33985] antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Tubules (Kidney))
Specification
Tubules (Kidney)
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate Buffer pH 6.0
Permeabilization
Yes - 0.2% Triton X-100
Blocking step
Serum 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 Dec 30 2010

Question

LOT NUMBER GR9270-1 ORDER NUMBER PO-11512 DESCRIPTION OF THE PROBLEM The antibody should stain mitochondria, but it actually stained no-specifically to all cell membrane. SAMPLE HeLa cells, BV2 cells, C8B4 cell and C6 cells PRIMARY ANTIBODY 1ug/ml as suggested by the data sheet (ie 1000x dilution). DETECTION METHOD Fluorescence microscope equipped with high sensitivity camera. POSITIVE AND NEGATIVE CONTROLS USED Positive control was other mitochondria antibody (ab5298 for HeLa cells only). Primary and 2nd antibodies only have been performed and none showed positive staining. ANTIBODY STORAGE CONDITIONS Aliqoted and stored at -20C FIXATION OF SAMPLE Paraformaldehyde (4%) for 15 min ANTIGEN RETRIEVAL n/a PERMEABILIZATION STEP Triton X-100 at 0.2% for 20 min (it work for other antibody) BLOCKING CONDITIONS 2% BSA for >30min SECONDARY ANTIBODY Anti-mouse IgG conjugated with Dylight 594 (ordered from Abcam #an97017). HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? >4 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? n/a ADDITIONAL NOTES the following message from the customer: "Please change to other abcam antibody that react with human, rat and mouse. The antibody should be raised from mouse. The antibody can be used for IF and WB. Would you please inform AbCam that I want to have ab14705 MS404 (Anti-MTCO1 antibody-Mitochondria Marker (mouse monoclonal) to replace my previous antibody (ab33985)?"

Read More

Abcam community

Verified customer

Asked on Sep 27 2011

Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody. The details you have kindly sent will provide us with vital information for our monitoring of product quality I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. As requested, we would be pleased to arrange this free of charge replacement. In order to do this, I am forwarding these details on to our distributors team who will be happy to organise this for you. Original order number: 940526 1 X vial ab33985 Anti-COX IV antibody [mAbcam33985 Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I can also recommend to consider the following suggestions for any future experiments, which I hope may help improve the results. Although other antibodies have worked well using this procedure, individual antibodies can sometimes require individual optimization. 1. I can recommend to try a lower dilution of antibody to reduce any non specific staining. Try 1:2000. 2. I can suggest to increase the concentration of blocking agent to 5% BSA 3. I can recommend to try including wash steps if you have not already done so. Try PBS containing 0.2% Tween (a more gentle detergent) 4 times for 5 minutes at each appropriate wash step. Thank you for your cooperation. Should you have any further questions, please do let me know. I would like to wish the customer best of luck with their future research.

Read More

Abcam Scientific Support

Answered on Sep 27 2011

1-10 of 26 Abreviews or Q&A

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