Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-LMAN1 antibody [EPR6979] (ab125006)

Reviews (2)Q&A (1)References (6)

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: ICC/IF, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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: ICC/IF, WB, IHC-Pmore details
    Unsuitable for: Flow Cyt or IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human LMAN1 aa 1-100. The exact sequence is proprietary.

  • Positive control

    • WB: HEK-293T, HeLa, SH-SY5Y, NCI-H460, Jurkat and JAR cell lysates. IHC-P: Human kidney tissue and Human papillary carcinoma of thyroid gland tissues.

  • 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.

Properties

Applications

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
ICC/IF 1/50 - 1/100.
WB 1/1000 - 1/10000. Detects a band of approximately 53 kDa (predicted molecular weight: 58 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. Heat up to 98 degrees C, below boiling, and then let cool for 10-20 min.
  • Application notes
    Is unsuitable for Flow Cyt or IP.

    • 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 : 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.

       

    • Immunocytochemistry/ Immunofluorescence - Anti-LMAN1 antibody [EPR6979] (ab125006)
      Immunocytochemistry/ Immunofluorescence - Anti-LMAN1 antibody [EPR6979] (ab125006)

      ICC/IF image of ab125006 stained HeLa cells. The cells were 100% methanol fixed (5 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 ab125006 at 1/100 dilution overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

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

      ab125006 at 1/100 dilution staining LMAN1 in paraffin-embedded Human kidney tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • 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 : HeLa cell lysate
      Lane 2 : 293T cell lysate
      Lane 3 : SH-SY5Y cell lysate
      Lane 4 : NCI-H460 cell lysate
      Lane 5 : Jurkat cell lysate
      Lane 6 : JAR cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit HRP at 1/2000 dilution

      Predicted band size: 58 kDa

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

      ab125006 at 1/100 dilution staining LMAN1 in paraffin-embedded Human papillary carcinoma of thyroid gland tissue by Immunohistochemistry.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    References

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

    ab125006 has been referenced in 6 publications.

    • Fu YL  et al. LMAN1 (ERGIC-53) promotes trafficking of neuroreceptors. Biochem Biophys Res Commun 511:356-362 (2019). PubMed: 30791981
    • 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
    • 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

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    1-3 of 3 Abreviews or Q&A

    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

    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

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