Recombinant Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)
![Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)](https://www.abcam.com/ps/products/197/ab197595/Images/ab197595-389452-anti-golph4gpp130-antibody-epr13439-western-blot-human-knockout-lysate.jpg "Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR13439] to GOLPH4/GPP130
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-GOLPH4/GPP130 antibody [EPR13439]
See all GOLPH4/GPP130 primary antibodies -
Description
Rabbit monoclonal [EPR13439] to GOLPH4/GPP130 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, HEK-293T and HepG2 whole cell lysates. IHC-P: Human renal adenocarcinoma tissue.
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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
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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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR13439 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab197595 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 |
|---|---|---|
| WB |
1/5000. Detects a band of approximately 130 kDa (predicted molecular weight: 82 kDa).
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| IHC-P |
1/500 - 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| Notes |
|---|
|
WB
1/5000. Detects a band of approximately 130 kDa (predicted molecular weight: 82 kDa). |
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IHC-P
1/500 - 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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Function
Plays a role in endosome to Golgi protein trafficking; mediates protein transport along the late endosome-bypass pathway from the early endosome to the Golgi. -
Sequence similarities
Belongs to the GOLIM4 family. -
Post-translational
modificationsPhosphorylated probably by c-AMP-dependent kinases in its lumenal part.
O-glycosylated; modified by sialic acid residues.
N-glycosylated; N-glycans are probably of the complex type and modified by sialic acid residues. -
Cellular localization
Golgi apparatus > Golgi stack membrane. Endosome membrane. Localizes to cis and medial Golgi cisternae. Probably cycles between early Golgi and distal compartments like endosome. - Information by UniProt
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Database links
- Entrez Gene: 27333 Human
- Omim: 606805 Human
- SwissProt: O00461 Human
- Unigene: 143600 Human
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Alternative names
- 130 kDa golgi localized phosphoprotein antibody
- cis antibody
- cis Golgi localized calcium binding protein antibody
see all
Images
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Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)All lanes : Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : GOLIM4 knockout HEK293T cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : MOLT-4 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 82 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab197595 observed at 130 kDa. Red - loading control ab8245 observed at 36 kDa.
ab197595 Anti-GOLPH4/GPP130 antibody [EPR13439] was shown to specifically react with GOLPH4/GPP130 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266123 (knockout cell lysate ab257455) was used. Wild-type and GOLPH4/GPP130 knockout samples were subjected to SDS-PAGE. ab197595 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)](https://www.abcam.com/ps/products/197/ab197595/Images/ab197595-237149-anti-golph4-antibody-epr13439-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)Immunohistochemical analysis of paraffin-embedded Human renal adenocarcinoma tissue labeling GOLPH4/GPP130 with ab197595 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Cytoplasm staining on Human renal adenocarcinoma tissue is observed. Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)](https://www.abcam.com/ps/products/197/ab197595/Images/ab197595-237151-anti-golph4-antibody-epr13439-western-blot.jpg)
Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595) at 1/2500 dilution + HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 82 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteBlocking/Dilution buffer: 5% NFDM/TBST.
The observed MW is consistent with what has been described in the literature PMID:20130081 and PMID:9201717
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![Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)](https://www.abcam.com/ps/products/197/ab197595/Images/ab197595-237150-anti-golph4-antibody-epr13439-western-blot.jpg)
Western blot - Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)All lanes : Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595) at 1/5000 dilution
Lane 1 : HepG2(Human liver hepatocellular carcinoma) whole cell lysate
Lane 2 : 293(Human embryonic kidney) whole cell lysate
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: 82 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The observed MW is consistent with what has been described in the literature (PMID:)20130081 and 9201717
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![Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)](https://www.abcam.com/ps/products/197/ab197595/Images/ab197595-5-benefits-of-recombinant-antibodies.png)
Anti-GOLPH4/GPP130 antibody [EPR13439] (ab197595)
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (1)
ab197595 has been referenced in 1 publication.
- Bonsergent E & Lavieu G Content release of extracellular vesicles in a cell-free extract. FEBS Lett 593:1983-1992 (2019). PubMed: 31175663
