Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-LMAN1 antibody [EPR6979] (ab125006)

Reviews (3)Q&A (1)References (7)
Western blot - Anti-LMAN1 antibody [EPR6979] (ab125006)
  • Immunocytochemistry - Anti-LMAN1 antibody [EPR6979] (ab125006)
  • Western blot - Anti-LMAN1 antibody [EPR6979] (ab125006)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LMAN1 antibody [EPR6979] (ab125006)
  • Flow Cytometry (Intracellular) - Anti-LMAN1 antibody [EPR6979] (ab125006)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LMAN1 antibody [EPR6979] (ab125006)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LMAN1 antibody [EPR6979] (ab125006)
  • 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
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • 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, ICCmore 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: 293T cells; IHC-P: Human, rat, and mouse kidney tissue sections. Flow Cyt (intra): 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • 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...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR6979

  • Isotype

    IgG

  • Research areas

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
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB (1)
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
1/50 - 1/100.
Notes
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
1/50 - 1/100.

Target

  • 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
    modifications

    The N-terminal may be partly blocked.

  • Cellular localization

    Endoplasmic reticulum-Golgi intermediate compartment membrane. Golgi apparatus membrane. Endoplasmic reticulum membrane.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • Endoplasmic reticulum golgi intermediate compartment protein 53 antibody
    • ER-Golgi intermediate compartment 53 kDa protein antibody
    • ERGIC-53 antibody
    • ERGIC53 antibody
    • ERGIC53 like protein antibody
    • F5F8D antibody
    • FMFD1 antibody
    • Gp58 antibody
    • Intracellular mannose specific lectin antibody
    • Intracellular mannose-specific lectin MR60 antibody
    • Lectin mannose binding 1 antibody
    • Lectin mannose-binding 1 antibody
    • Lman1 antibody
    • LMAN1 like protein antibody
    • LMAN1_HUMAN antibody
    • MCFD1 antibody
    • MR60 antibody
    • Protein ERGIC-53 antibody
    see all

Images

  • Western blot - Anti-LMAN1 antibody [EPR6979] (ab125006)
    Western blot - Anti-LMAN1 antibody [EPR6979] (ab125006)
    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 - Anti-LMAN1 antibody [EPR6979] (ab125006)
    Immunocytochemistry - Anti-LMAN1 antibody [EPR6979] (ab125006)
    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.
  • Western blot - Anti-LMAN1 antibody [EPR6979] (ab125006)
    Western blot - Anti-LMAN1 antibody [EPR6979] (ab125006)
    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.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LMAN1 antibody [EPR6979] (ab125006)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LMAN1 antibody [EPR6979] (ab125006)
    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.
  • Flow Cytometry (Intracellular) - Anti-LMAN1 antibody [EPR6979] (ab125006)
    Flow Cytometry (Intracellular) - Anti-LMAN1 antibody [EPR6979] (ab125006)

    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).

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LMAN1 antibody [EPR6979] (ab125006)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LMAN1 antibody [EPR6979] (ab125006)
    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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LMAN1 antibody [EPR6979] (ab125006)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LMAN1 antibody [EPR6979] (ab125006)
    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.
  • Anti-LMAN1 antibody [EPR6979] (ab125006)
    Anti-LMAN1 antibody [EPR6979] (ab125006)

References (7)

Publishing research using ab125006? Please let us know so that we can cite the reference in this datasheet.

ab125006 has been referenced in 7 publications.

  • Camus SM  et al. CHC22 clathrin mediates traffic from early secretory compartments for human GLUT4 pathway biogenesis. J Cell Biol 219:N/A (2020). PubMed: 31863584
  • Bassaganyas L  et al. New factors for protein transport identified by a genome-wide CRISPRi screen in mammalian cells. J Cell Biol 218:3861-3879 (2019). PubMed: 31488582
  • Meier S  et al. The molecular basis of chaperone-mediated interleukin 23 assembly control. Nat Commun 10:4121 (2019). PubMed: 31511508
  • Fu YL  et al. LMAN1 (ERGIC-53) promotes trafficking of neuroreceptors. Biochem Biophys Res Commun 511:356-362 (2019). PubMed: 30791981
  • Emmer BT  et al. The cargo receptor SURF4 promotes the efficient cellular secretion of PCSK9. Elife 7:N/A (2018). PubMed: 30251625
  • Fan J  et al. cTAGE5 deletion in pancreatic ß cells impairs proinsulin trafficking and insulin biogenesis in mice. J Cell Biol 216:4153-4164 (2017). PubMed: 29133483
  • Duellman T  et al. LMAN1 (ERGIC-53) is a potential carrier protein for matrix metalloproteinase-9 glycoprotein secretion. Biochem Biophys Res Commun 464:685-91 (2015). ICC/IF ; Human . PubMed: 26150355

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-4 of 4 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-LMAN1 antibody [EPR6979]

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Rat hepatocytes)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
Rat hepatocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Read More

Dr. Armen Petrosyan

Verified customer

Submitted Sep 10 2020

Western blot abreview for Anti-LMAN1 antibody [EPR6979]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (22Rv1 - prostate cancer cells)
Gel Running Conditions
Reduced Denaturing (8% SDS-PAGE)
Loading amount
20 µg
Specification
22Rv1 - prostate cancer cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More

Dr. Armen Petrosyan

Verified customer

Submitted Jul 18 2019

Immunocytochemistry/ Immunofluorescence abreview for Anti-LMAN1 antibody [EPR6979]

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (RWPE-1 - prosate epithelial cells)
Permeabilization
Yes - 0.2% Triton X-100
Specification
RWPE-1 - prosate epithelial cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Read More

Dr. Armen Petrosyan

Verified customer

Submitted Jul 18 2019

Question

What are the immunogen sequences for the LMAN1 antibodies ab125006 and ab126720? Do the immunogens overlap with the various binding sites between amino acids 88 - 181?
What cells were tested in ICC? Do you have any images of this testing?

Read More
Answer

Thank you for contacting us. The specific immunogen sequences for these antibodies are proprietary. They are both internal sequences, but do not overlap with amino acids 88 - 181.
The lab has tested HeLa and JAR cells in immunocytochemistry, but unfortunately we do not have any images from this testing. I hope this helps, please let me know if you need any additional information or assistance.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com