Recombinant 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
Related conjugates and formulations
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
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
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
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. |
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
- 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
see all -
Alternative names
- 70 kDa peroxisomal membrane protein antibody
- ABC 43 antibody
- ABC D3 antibody
see all
Images
-
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 kDaLanes 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.
-
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).
-
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 kDaLanes 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.
-
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? -
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).
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (6)
ab109448 has been referenced in 6 publications.
- Luo F et al. Hepatic TM6SF2 Is Required for Lipidation of VLDL in a Pre-Golgi Compartment in Mice and Rats. Cell Mol Gastroenterol Hepatol 13:879-899 (2022). PubMed: 34923175
- Chang HC et al. The BAX-binding protein MOAP1 associates with LC3 and promotes closure of the phagophore. Autophagy 17:3725-3739 (2021). PubMed: 33783314
- 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