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RecombinantRabMAb

Recombinant Anti-HGFA Inhibitor 2 antibody [EPR6283(2)] (ab128926)

Submit a review Q&A (2)References (3)
Western blot - Anti-HGFA Inhibitor 2 antibody [EPR6283(2)] (ab128926)
  • Immunocytochemistry/ Immunofluorescence - Anti-HGFA Inhibitor 2 antibody [EPR6283(2)] (ab128926)
  • OI-RD Scanning - Anti-HGFA Inhibitor 2 antibody [EPR6283(2)] (ab128926)
  • Anti-HGFA Inhibitor 2 antibody [EPR6283(2)] (ab128926)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6283(2)] to HGFA Inhibitor 2
  • Suitable for: WB, ICC/IF
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-HGFA Inhibitor 2 antibody [EPR6283(2)]
    See all HGFA Inhibitor 2 primary antibodies

  • Description

    Rabbit monoclonal [EPR6283(2)] to HGFA Inhibitor 2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IFmore details
    Unsuitable for: IHC-P or IP

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat

  • Immunogen

    Synthetic peptide within Human HGFA Inhibitor 2 aa 100-200 (extracellular). The exact sequence is proprietary.

  • Positive control

    • WB: Human Caco-2, MCF7, PC-3, BxPC-3, MDA-MB-435S whole cell lysates.

  • 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

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab128926 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
1/1000 - 1/10000. Detects a band of approximately 34 kDa (predicted molecular weight: 28 kDa).
ICC/IF
1/250 - 1/500.
Notes
WB
1/1000 - 1/10000. Detects a band of approximately 34 kDa (predicted molecular weight: 28 kDa).
ICC/IF
1/250 - 1/500.
Application notes
Is unsuitable for IHC-P or IP.

Target

  • Function

    Inhibitor of HGF activator. Also inhibits plasmin, plasma and tissue kallikrein, and factor XIa.

  • Tissue specificity

    Expressed in placenta, kidney, pancreas, prostate, testis, thymus, and trachea.

  • Involvement in disease

    Defects in SPINT2 are the cause of diarrhea type 3 (DIAR3) [MIM:270420]; also known as congenital sodium diarrhea (CSD). DIAR3 is a rare, inherited diarrhea of infancy. A diagnosis of DIAR3 is made on the findings of a life-threatening secretory diarrhea, severe metabolic acidosis, and hyponatremia secondary to extraordinarily high fecal losses of sodium, with low or normal excretion of urinary sodium, in the absence of infectious, autoimmune, and endocrine causes.

  • Sequence similarities

    Contains 2 BPTI/Kunitz inhibitor domains.

  • Domain

    This inhibitor contains two inhibitory domains.

  • Cellular localization

    Membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • AL024025 antibody
      • Bikunin, placental antibody
      • C76321 antibody
      • DIAR3 antibody
      • FLJ45571 antibody
      • HAI-2 antibody
      • HAI2 antibody
      • Hepatocyte growth factor activator inhibitor type 2 antibody
      • Kop antibody
      • Kunitz type protease inhibitor 2 antibody
      • Kunitz-type protease inhibitor 2 antibody
      • Kunitz-type serine protease inhibitor antibody
      • MGC72638 antibody
      • PB antibody
      • Placental bikunin antibody
      • Serine peptidase inhibitor Kunitz type 2 antibody
      • Serine protease inhibitor, Kunitz type, 2 antibody
      • SPINT2 antibody
      • SPIT2_HUMAN antibody
      see all

    Images

    • Western blot - Anti-HGFA Inhibitor 2 antibody [EPR6283(2)] (ab128926)
      Western blot - Anti-HGFA Inhibitor 2 antibody [EPR6283(2)] (ab128926)
      All lanes : Anti-HGFA Inhibitor 2 antibody [EPR6283(2)] (ab128926) at 1/1000 dilution

      Lane 1 : Caco-2 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate at 15 µg
      Lane 2 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg
      Lane 3 : PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg
      Lane 4 : BxPC-3 (Human pancreas adenocarcinoma epithelial cell) whole cell lysate at 20 µg
      Lane 5 : MDA-MB-435S (Human mammary gland ductal carcinoma melanocyte) whole cell lysate at 20 µg

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 28 kDa
      Observed band size: 34 kDa why is the actual band size different from the predicted?



      Blocking buffer: 5% NFDM/TBST.

    • Immunocytochemistry/ Immunofluorescence - Anti-HGFA Inhibitor 2 antibody [EPR6283(2)] (ab128926)
      Immunocytochemistry/ Immunofluorescence - Anti-HGFA Inhibitor 2 antibody [EPR6283(2)] (ab128926)
      ab128926, at 1/250 dilution, staining HGFA Inhibitor 2 in MCF-7 cells by Immunofluorescence.
    • OI-RD Scanning - Anti-HGFA Inhibitor 2 antibody [EPR6283(2)] (ab128926)
      OI-RD Scanning - Anti-HGFA Inhibitor 2 antibody [EPR6283(2)] (ab128926)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-HGFA Inhibitor 2 antibody [EPR6283(2)] (ab128926)
      Anti-HGFA Inhibitor 2 antibody [EPR6283(2)] (ab128926)

    References (3)

    Publishing research using ab128926? Please let us know so that we can cite the reference in this datasheet.

    ab128926 has been referenced in 3 publications.

    • Ma Z  et al. STYK1 promotes tumor growth and metastasis by reducing SPINT2/HAI-2 expression in non-small cell lung cancer. Cell Death Dis 10:435 (2019). PubMed: 31164631
    • Roversi FM  et al. Up-regulation of SPINT2/HAI-2 by Azacytidine in bone marrow mesenchymal stromal cells affects leukemic stem cell survival and adhesion. J Cell Mol Med N/A:N/A (2018). PubMed: 30484958
    • Roversi FM  et al. Serine protease inhibitor kunitz-type 2 is downregulated in myelodysplastic syndromes and modulates cell-cell adhesion. Stem Cells Dev 23:1109-20 (2014). PubMed: 24410667

    Customer reviews and Q&As

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

    1-2 of 2 Abreviews or Q&A

    Question

    Hi,

    I was wondering if the cell lysate sample you have sent me requires a boiling step prior to use. It doesn't seem to be mentioned in the data sheet of the cell lysate. I have run the sample and performed an immunoblot for SPINT2, however it was negative as well. THP-1 did have a band right below 37kDa for ab128926. ab83056 did not work at all.

    Thank you,

    Read More
    Answer

    Thank you for contacting Abcam.

    Listed in the protocol book that I sent you is the western blot protocol we recommend. Since the HGFA Inhibitor 2 protein is located on the cell membrane, we recommend using a NP-40 or RIPA buffer for cell lysis. Additionally, protease and phosphatase inhibitors should be added fresh to the lysis buffer for each preparation.

    As stated in the protocol, the samples need to be denatured and reduced. Denaturing is done by boiling in SDS at 95-100oC for 5 minutes.

    I've attached the protocol book again, which explains these steps in detail as well as troubleshooting options.

    Here is a link to our western blot guide as well:
    https://www.abcam.com/ps/pdf/protocols/WB-beginner.pdf

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

    Read More

    Question

    ab128926:
    Application: WB
    Positive controls run: MCF7 cell lysate (listed on datasheet, did not work), THP cell lysate (not listed as positive control on datasheet, worked)
    Immunogen: external
    Storage: -20C upon arrival
    ab83056:
    Application: WB
    Positive controls run: THP cell lysate (gives multiple bands, nothing in predicted / expected range of 25-34 kDa)
    Protocol:
    5% milk block + TBS-T, 1 hr, RT
    Primary: 1:1000 O/N 4C
    Immunogen: internal
    Storage: Aliquoted upon arrival
    Sample: 10ug / lane for positive control, 40ug / lane for experimentals
    Buffer: RIPA

    Read More
    Answer

    Thank you for contacting Abcam.

    Forwarded below is the information that you told me over the phone regarding ab128926 and ab83056. You mentioned ab128926 did produce a positive signal in the THP cell lysate while ab83056 did not produce a positive signal in the same THP cell lysate. As we discussed, I am sending you a positive control free of charge (Order XXX, ab7914 HL60 whole cell lysate). I expect the whole cell lysate to contain the HGFA Inhibitor 2 protein based on the information I obtained from protein atlas in the following link:
    http://www.proteinatlas.org/ENSG00000167642/cell

    We also discussed some troubleshooting steps to the western blot protocol you were using, specifically for ab83056 due to the presence of non-specific bands and lack of positive bands, listed below:
    1. Lower primary antibody concentration
    2. Load 20ug of positive control rather than 10ug
    3. Block for longer than 1 hour at room temperature using 3% BSA rather than 5% milk

    I also attached our protocol book, which provides further suggestions for troubleshooting when non-specific bands are observed in western blot.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

    Read More

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