Recombinant Anti-LMAN1 antibody [EPR6979] (ab125006)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6979] to LMAN1
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-LMAN1 antibody [EPR6979]
See all LMAN1 primary antibodies -
Description
Rabbit monoclonal [EPR6979] to LMAN1 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human LMAN1 aa 1-100. The exact sequence is proprietary.
Database link: P49257 -
Positive control
- WB: Jurkat, HEK-293T and HeLa cell lysates; Mouse and Rat spleen and heart lysates; ICC/IF: 293T cells; IHC-P: Human, rat, and mouse kidney tissue sections. Flow Cyt (intra): HeLa cells.
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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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6979 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab125006 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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB | (1) |
1/1000 - 1/10000. Detects a band of approximately 53 kDa (predicted molecular weight: 58 kDa).
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IHC-P |
1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. Heat up to 98°C, below boiling, and then let cool for 10-20 min. For unpurified use at 1/100 - 1/250. |
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ICC/IF | (3) |
1/50 - 1/100.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
1/1000 - 1/10000. Detects a band of approximately 53 kDa (predicted molecular weight: 58 kDa). |
IHC-P
1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. Heat up to 98°C, below boiling, and then let cool for 10-20 min. For unpurified use at 1/100 - 1/250. |
ICC/IF
1/50 - 1/100. |
Target
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Function
Mannose-specific lectin. May recognize sugar residues of glycoproteins, glycolipids, or glycosylphosphatidyl inositol anchors and may be involved in the sorting or recycling of proteins, lipids, or both. The LMAN1-MCFD2 complex forms a specific cargo receptor for the ER-to-Golgi transport of selected proteins. -
Tissue specificity
Ubiquitous. -
Involvement in disease
Defects in LMAN1 are THE cause of factor V and factor VIII combined deficiency type 1 (F5F8D1) [MIM:227300]; also known as multiple coagulation factor deficiency I (MCFD1). F5F8D1 is an autosomal recessive blood coagulation disorder characterized by bleeding symptoms similar to those in hemophilia or parahemophilia, that are caused by single deficiency of FV or FVIII, respectively. The most common symptoms are epistaxis, menorrhagia, and excessive bleeding during or after trauma. Plasma levels of coagulation factors V and VIII are in the range of 5 to 30% of normal. -
Sequence similarities
Contains 1 L-type lectin-like domain. -
Post-translational
modificationsThe N-terminal may be partly blocked. -
Cellular localization
Endoplasmic reticulum-Golgi intermediate compartment membrane. Golgi apparatus membrane. Endoplasmic reticulum membrane. - Information by UniProt
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Database links
- Entrez Gene: 3998 Human
- Entrez Gene: 70361 Mouse
- Entrez Gene: 116666 Rat
- Omim: 601567 Human
- SwissProt: P49257 Human
- SwissProt: Q9D0F3 Mouse
- SwissProt: Q62902 Rat
- Unigene: 465295 Human
see all -
Alternative names
- Endoplasmic reticulum golgi intermediate compartment protein 53 antibody
- ER-Golgi intermediate compartment 53 kDa protein antibody
- ERGIC-53 antibody
see all
Images
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All lanes : Anti-LMAN1 antibody [EPR6979] (ab125006) at 1/1000 dilution
Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Mouse spleen lysate
Lane 4 : Mouse heart lysate
Lane 5 : Rat spleen lysate
Lane 6 : Rat heart lysate
Lane 7 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 58 kDa -
Immunocytochemistry analysis of 293T (Human embryonic kidney epithelial cell) cells labeling LMAN1 with Purified ab125006 at 1/50 dilution (2 µg/mL). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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All lanes : Anti-LMAN1 antibody [EPR6979] (ab125006) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : LMAN1 knockout HEK-293T cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Daudi cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 58 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab125006 observed at 55 kDa. Red - loading control ab8245 observed at 37 kDa.
ab125006 Anti-LMAN1 antibody [EPR6979] was shown to specifically react with Protein ERGIC-53 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266248 (knockout cell lysate ab257505) was used. Wild-type and Protein ERGIC-53 knockout samples were subjected to SDS-PAGE. ab125006 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°CC 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) analysis of human kidney tissue sections labeling LMAN1 with Purified ab125006 at 1/500 dilution (0.21 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling LMAN1 with Purified ab125006 at 1/20 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling LMAN1 with Purified ab125006 at 1/500 dilution (0.21 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling LMAN1 with Purified ab125006 at 1/500 dilution (0.21 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (14)
ab125006 has been referenced in 14 publications.
- Devireddy S & Ferguson SM Efficient progranulin exit from the ER requires its interaction with prosaposin, a Surf4 cargo. J Cell Biol 221:N/A (2022). PubMed: 34919127
- 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
- Chen D et al. Angiogenesis depends upon EPHB4-mediated export of collagen IV from vascular endothelial cells. JCI Insight 7:N/A (2022). PubMed: 35015735
- Wang M et al. Pharmacological activation of ATF6 remodels the proteostasis network to rescue pathogenic GABAA receptors. Cell Biosci 12:48 (2022). PubMed: 35477478
- Fan BY et al. Liproxstatin-1 is an effective inhibitor of oligodendrocyte ferroptosis induced by inhibition of glutathione peroxidase 4. Neural Regen Res 16:561-566 (2021). PubMed: 32985488