Validated using a knockout cell line

Anti-PMP70 antibody (ab85550)

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Western blot - Anti-PMP70 antibody (ab85550)
  • Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab85550)
  • Western blot - Anti-PMP70 antibody (ab85550)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMP70 antibody (ab85550)
  • Western blot - Anti-PMP70 antibody (ab85550)

Key features and details

  • Rabbit polyclonal to PMP70
  • Suitable for: IHC-P, ICC/IF, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

Overview

  • Product name

    Anti-PMP70 antibody
    See all PMP70 primary antibodies

  • Description

    Rabbit polyclonal to PMP70

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Cow, Orangutan

  • Immunogen

    Synthetic peptide corresponding to Rat PMP70 aa 600 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab94821)

  • Positive control

    • This antibody gave a positive signal in the following tissue lysates: Rat liver; Rat kidney; Human liver; Mouse Lung; as well as the following whole cell lysates: HepG2; HeLa.

  • General notes

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab85550 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
IHC-P
Use a concentration of 1 µg/ml.
ICC/IF
Use a concentration of 1 µg/ml.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 73 kDa (predicted molecular weight: 75 kDa).
Notes
IHC-P
Use a concentration of 1 µg/ml.
ICC/IF
Use a concentration of 1 µg/ml.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 73 kDa (predicted molecular weight: 75 kDa).

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.
  • Information by UniProt

  • Database links

    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 (ab85550)
    Western blot - Anti-PMP70 antibody (ab85550)
    All lanes : Anti-PMP70 antibody (ab85550) at 1 µg/ml

    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 - ab85550 observed at 75 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab85550 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. Ab85550 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab85550)
    Immunocytochemistry/ Immunofluorescence - Anti-PMP70 antibody (ab85550)
    ICC/IF image of ab85550 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab85550, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HepG2 cells at 1µg/ml.
  • Western blot - Anti-PMP70 antibody (ab85550)
    Western blot - Anti-PMP70 antibody (ab85550)
    All lanes : Anti-PMP70 antibody (ab85550) at 1 µg/ml

    Lane 1 : Liver (Rat) Tissue Lysate
    Lane 2 : Kidney (Rat) Tissue Lysate
    Lane 3 : Human liver tissue lysate - total protein (ab29889)
    Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 5 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

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

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 75 kDa
    Observed band size: 70 kDa why is the actual band size different from the predicted?
    Additional bands at: 25 kDa, 28 kDa, 37 kDa, 50 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 2 minutes


    The 73 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to Rat ATP-binding cassette sub-family D member 3 (PMP70).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMP70 antibody (ab85550)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMP70 antibody (ab85550)
    IHC image of Ab85550 staining in Human Cervical carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with Ab85550, 1µg/ml, for 15 mins 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.
  • Western blot - Anti-PMP70 antibody (ab85550)
    Western blot - Anti-PMP70 antibody (ab85550)
    Anti-PMP70 antibody (ab85550) at 1 µg/ml + Lung (Mouse) Tissue Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

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


    Exposure time: 1 minute

References (5)

Publishing research using ab85550? Please let us know so that we can cite the reference in this datasheet.

ab85550 has been referenced in 5 publications.

  • Demaret T  et al. Longitudinal study of Pex1-G844D NMRI mouse model: A robust pre-clinical model for mild Zellweger spectrum disorder. Biochim Biophys Acta Mol Basis Dis 1866:165900 (2020). PubMed: 32693164
  • Elliott EI  et al. Cutting Edge: Mitochondrial Assembly of the NLRP3 Inflammasome Complex Is Initiated at Priming. J Immunol 200:3047-3052 (2018). PubMed: 29602772
  • Ohoka N  et al. SNIPER(TACC3) induces cytoplasmic vacuolization and sensitizes cancer cells to Bortezomib. Cancer Sci 108:1032-1041 (2017). PubMed: 28192613
  • Sidjanin DJ  et al. TBC1D20 mediates autophagy as a key regulator of autophagosome maturation. Autophagy 12:1759-1775 (2016). PubMed: 27487390
  • Giordano CR  et al. Amyloid-beta neuroprotection mediated by a targeted antioxidant. Sci Rep 4:4983 (2014). ICC/IF ; Rat . PubMed: 24828380

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