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    products/primary-antibodies/pmp70-antibody-epr5614-ab109448.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-PMP70 antibody [EPR5614] (ab109448)

  • Datasheet
  • SDS
Reviews (3) Submit a question References (4)

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Western blot - Anti-PMP70 antibody [EPR5614] (ab109448)
  • Flow Cytometry (Intracellular) - Anti-PMP70 antibody [EPR5614] (ab109448)
  • Western blot - Anti-PMP70 antibody [EPR5614] (ab109448)
  • Western blot - Anti-PMP70 antibody [EPR5614] (ab109448)
  • Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody [EPR5614] (ab109448)
  • OI-RD Scanning - Anti-PMP70 antibody [EPR5614] (ab109448)
  • Anti-PMP70 antibody [EPR5614] (ab109448)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5614] to PMP70
  • Suitable for: Flow Cyt (Intra), WB, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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Overview

  • Product name

    Anti-PMP70 antibody [EPR5614]
    See all PMP70 primary antibodies
  • Description

    Rabbit monoclonal [EPR5614] to PMP70
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

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

  • Positive control

    • WB: Hap1, HeLa, 293T, SH SY5Y, Caco2, and HepG2 cell lysates. Flow Cyt (intra): Permeabilized HepG2 cells. ICC : Permeabilized HeLa 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 -20°C. Stable for 12 months at -20°C.
  • Dissociation constant (KD)

    KD = 4.10 x 10 -11 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR5614
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Other
    • Tags & Cell Markers
    • Subcellular Markers
    • Organelles
    • Peroxisome
    • Signal Transduction
    • Protein Trafficking
    • Organelle Proteins
    • Signal Transduction
    • Metabolism
    • Lipid metabolism
    • Metabolism
    • Types of disease
    • Neurodegenerative disease
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-PMP70 antibody [EPR5614] (ab199018)
    • Alexa Fluor® 647 Anti-PMP70 antibody [EPR5614] (ab199019)
    • Anti-PMP70 antibody [EPR5614] - BSA and Azide free (ab247868)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab109448 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)
1/100 - 1/500.

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

WB (3)
1/1000 - 1/10000. Detects a band of approximately 70 kDa (predicted molecular weight: 75 kDa).
ICC/IF
1/250.
Notes
Flow Cyt (Intra)
1/100 - 1/500.

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

WB
1/1000 - 1/10000. Detects a band of approximately 70 kDa (predicted molecular weight: 75 kDa).
ICC/IF
1/250.
Application notes
Is unsuitable for IHC-P or IP.

Target

  • Function

    Probable transporter. The nucleotide-binding fold acts as an ATP-binding subunit with ATPase activity.
  • Sequence similarities

    Belongs to the ABC transporter superfamily. ABCD family. Peroxisomal fatty acyl CoA transporter (TC 3.A.1.203) subfamily.
    Contains 1 ABC transmembrane type-1 domain.
    Contains 1 ABC transporter domain.
  • Cellular localization

    Peroxisome membrane.
  • Target information above from: UniProt accession P28288 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5825 Human
    • Entrez Gene: 19299 Mouse
    • Entrez Gene: 25270 Rat
    • Omim: 170995 Human
    • SwissProt: P28288 Human
    • SwissProt: P55096 Mouse
    • SwissProt: P16970 Rat
    • Unigene: 700576 Human
    • Unigene: 399042 Mouse
    • Unigene: 7024 Rat
    see all
  • Alternative names

    • 70 kDa peroxisomal membrane protein antibody
    • ABC 43 antibody
    • ABC D3 antibody
    • ABC43 antibody
    • ABCD 3 antibody
    • Abcd3 antibody
    • ABCD3 protein antibody
    • ABCD3_HUMAN antibody
    • ATP binding cassette sub family D (ALD) member 3 antibody
    • ATP binding cassette sub family D member 3 antibody
    • ATP-binding cassette sub-family D member 3 antibody
    • CBAS5 antibody
    • dJ824O18.1 (ATP-binding cassette, sub family D (ALD), member 3 (PMP70, PXMP1)) antibody
    • Peroxisomal membrane protein 1 (70kD) antibody
    • Peroxisomal membrane protein 1 (70kD, Zellweger syndrome) antibody
    • Peroxisomal membrane protein 1 antibody
    • PMP 70 antibody
    • PMP70 antibody
    • PXMP 1 antibody
    • PXMP1 antibody
    • ZWS2 antibody
    see all

Images

  • Western blot - Anti-PMP70 antibody [EPR5614] (ab109448)
    Western blot - Anti-PMP70 antibody [EPR5614] (ab109448)
    All lanes : Anti-PMP70 antibody [EPR5614] (ab109448) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : ABCD3 knockout HeLa cell lysate

    Lysates/proteins at 40 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 75 kDa
    Observed band size: 75 kDa



    Lanes 1- 2: Merged signal (red and green). Green - ab109448 observed at 75 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab109448 was shown to react with PMP70 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265294 (knockout cell lysate ab257134) was used. Wild-type HeLa and ABCD3 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109448 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry (Intracellular) - Anti-PMP70 antibody [EPR5614] (ab109448)
    Flow Cytometry (Intracellular) - Anti-PMP70 antibody [EPR5614] (ab109448)

    ab109448 at 1/100 dilution staining PMP70 in permeabilized HepG2 cells by intracellular flow cytometry (shown in red). Rabbit IgG negative control (shown in green).

     

  • Western blot - Anti-PMP70 antibody [EPR5614] (ab109448)
    Western blot - Anti-PMP70 antibody [EPR5614] (ab109448)
    All lanes : Anti-PMP70 antibody [EPR5614] (ab109448) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : ABCD3 (PMP70) knockout HAP1 whole cell lysate
    Lane 3 : A431 whole cell lysate
    Lane 4 : HEPG2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 75 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab109448 observed at 75 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab109448 was shown to specifically react with PMP70 in wild-type HAP1 cells as signal was lost in ABCD3 (PMP70) knockout cells. Wild-type and ABCD3 (PMP70) knockout samples were subjected to SDS-PAGE. Ab109448 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-PMP70 antibody [EPR5614] (ab109448)
    Western blot - Anti-PMP70 antibody [EPR5614] (ab109448)
    All lanes : Anti-PMP70 antibody [EPR5614] (ab109448) at 1/1000 dilution

    Lane 1 : 293T cell lysate
    Lane 2 : SH SY5Y cell lysate
    Lane 3 : Caco2 cell lysate
    Lane 4 : HepG2 cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 75 kDa
    Observed band size: 70 kDa why is the actual band size different from the predicted?

  • Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody [EPR5614] (ab109448)
    Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody [EPR5614] (ab109448)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling PMP70 with primary antibody anti-PMP70 (ab109448) at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150081) secondary antibody at 1/1000 dilution (2.0 µg/mL). Confocal image showing cytoplasmic staining on HeLa cells. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL). The nuclear counter stain is DAPI (blue).
    The secondary antibody only control is : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (2.0 µg/mL).

  • OI-RD Scanning - Anti-PMP70 antibody [EPR5614] (ab109448)
    OI-RD Scanning - Anti-PMP70 antibody [EPR5614] (ab109448)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Anti-PMP70 antibody [EPR5614] (ab109448)
    Anti-PMP70 antibody [EPR5614] (ab109448)

Protocols

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

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (4)

Publishing research using ab109448? Please let us know so that we can cite the reference in this datasheet.

ab109448 has been referenced in 4 publications.

  • Covill-Cooke C  et al. Peroxisomal fission is modulated by the mitochondrial Rho-GTPases, Miro1 and Miro2. EMBO Rep 21:e49865 (2020). PubMed: 31894645
  • Torres SE  et al. Ceapins block the unfolded protein response sensor ATF6a by inducing a neomorphic inter-organelle tether. Elife 8:N/A (2019). PubMed: 31149896
  • Chang CL  et al. Spastin tethers lipid droplets to peroxisomes and directs fatty acid trafficking through ESCRT-III. J Cell Biol 218:2583-2599 (2019). PubMed: 31227594
  • Cai M  et al. Disruption of peroxisome function leads to metabolic stress, mTOR inhibition, and lethality in liver cancer cells. Cancer Lett 421:82-93 (2018). ICC/IF . PubMed: 29458144

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Western blot abreview for Anti-PMP70 antibody [EPR5614]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (kidney)
Gel Running Conditions
Reduced Denaturing (4-12% bis-tris gel, MES buffer)
Loading amount
30 µg
Specification
kidney
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 Nov 03 2020

Western blot abreview for Anti-PMP70 antibody [EPR5614]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Dopaminergic Neurons)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
Dopaminergic Neurons
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 Oct 19 2020

Western blot abreview for Anti-PMP70 antibody [EPR5614]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
20 µg
Specification
HEK293T
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · 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.

Abcam user community

Verified customer

Submitted Apr 29 2019

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