Anti-A proliferation-inducing ligand antibody [Sacha-1] (ab16085)

Overview

  • Product name
    Anti-A proliferation-inducing ligand antibody [Sacha-1]
    See all A proliferation-inducing ligand primary antibodies
  • Description
    Rat monoclonal [Sacha-1] to A proliferation-inducing ligand
  • Host species
    Rat
  • Tested applications
    Suitable for: Flow Cyt, ICCmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Fusion protein corresponding to Human A proliferation-inducing ligand aa 105-250. Fused to amino acids 16 - 108 of human headless, ACRP30.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity
    >95% by SDS-PAGE
  • Clonality
    Monoclonal
  • Clone number
    Sacha-1
  • Isotype
    IgG2a
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab16085 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 Use a concentration of 1 µg/ml.

ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

ICC Use a concentration of 5 µg/ml.

Target

  • Function
    Cytokine that binds to TNFRSF13B/TACI and to TNFRSF17/BCMA. May be implicated in the regulation of tumor cell growth. May be involved in monocyte/macrophage-mediated immunological processes.
  • Tissue specificity
    Expressed at high levels in transformed cell lines, cancers of colon, thyroid, lymphoid tissues and specifically expressed in monocytes and macrophages.
  • Sequence similarities
    Belongs to the tumor necrosis factor family.
  • Post-translational
    modifications
    The precursor is cleaved by furin.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • A proliferation inducing ligand antibody
    • A proliferation-inducing ligand antibody
    • APRIL antibody
    • CD256 antibody
    • TALL-2 antibody
    • TALL2 antibody
    • TNF related death ligand antibody
    • TNF- and APOL-related leukocyte expressed ligand 2 antibody
    • TNF-related death ligand 1 antibody
    • TNF13_HUMAN antibody
    • TNFSF13 antibody
    • TRDL-1 antibody
    • TRDL1 antibody
    • Tumor necrosis factor (ligand) superfamily, member 13 antibody
    • Tumor necrosis factor ligand superfamily member 13 antibody
    • Tumor necrosis factor like protein ZTNF2 antibody
    • Tumor necrosis factor related death ligand 1 antibody
    • UNQ383/PRO715 antibody
    • ZTNF2 antibody
    see all

Images

  • ab16085 at 1ug/ml detecting  membrane-bound human A proliferation-inducing ligand on HEK 293T cells by FACS analysis.

     

    HEK 293T cells (5x105) were mock transfected (thin line) or transfected with an expression plasmid enabling surface expression of a non-cleavable mouse A proliferation-inducing ligand (thick line). Cells were incubated on ice for 30 min. in 50ul FACS buffer (PBS, 5% fetal calf serum, 0.02% azide) containing 1ug/ml of MAb to APRIL (Sacha-1). After washing in FACS buffer, PE-conjugated antibody to rat IgG was added. Cells were incubated on ice for 30 min., washed and then analyzed by flow cytometry.

  • Immunostaining of HEK-293T cells transfected with a murine A proliferation-inducing ligand expression plasmid (left panels), or mock transfected (right panel).

    Three days after transfection of cells with the indicated constructs, cells were fixed with acetone. Slides were blocked with IgG, and incubated for 1 hour with 5ug/ml ab16085 or control murine IgG (isotype control) in 1%BSA / 1x PBS for 1 hour. After washes wish PBS, samples were incubated with the secondary Ab for 1 hour, washed in PBS and revealed with StreptABComplex/HRP (Vector) and AEC.

References

ab16085 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Thank you for your email. I am sorry for the delay as lab took more time than anticipated.

We may be able to provide you the positive control lysates for ab16088. This would be whole cell lysate of HEK 293T cells transfected with an expression plasmid for human APRIL (aa 1-250) fused at the C-terminus to a FLAG®-tag. We used the same lysate to test ab16088. However because ab16085 cross react with mouse and human so this antibody might also cross react.

Could you let me if you are still interested in buying this lysate? I would be able to provide you the catalogue number soon.

Once again thank you very much for having patience!

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Answer

Thank you for contacting us.

I can confirm that, I have sent reminder to our collaborator lab. I will send you the requested details as soon as I receive a reply from them.

Many thanks for having patience!

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Question

BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM no signal. We have tested the antibody on T cells (activated and not activated) as well as macrophages (resting and activated) and have not seen any staining above isotype. SAMPLE Murine macrophages and T cells. Human T cells. PRIMARY ANTIBODY Sacha-1/human,mouse/facs buffer/1:50/30 minutes, washed with perm wash buffer from IC kit DETECTION METHOD FACS and LSR POSITIVE AND NEGATIVE CONTROLS USED Positive controls for were T cells and macrophages (activated and resting) and negative controls were B cells as well as isotype controls. ANTIBODY STORAGE CONDITIONS 4 degrees SAMPLE PREPARATION Mouse splenocytes were ACK treated to purify lymphocytes. Human whole blood was ACK treated. NUMBER OF CELLS USED Roughly 2 X 10 7 cells per sample. PERMEABILIZATION STEP Cells were permeablized using e bioscience kit. And fixed after staining was complete. Antibody was also tested under non permabilizing conditions and without fixation. BLOCKING CONDITIONS 2% mouse serum, diluted in FACS buffer at 4 degrees for fifteen mintues. SECONDARY ANTIBODY PharMingen/rabbit(?)/facs buffer/ 1:50/30 min., wased with perm wash buffer from IC Kit HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? I have tried detecting APRIL staining in a higher expressing population, I have tried looking at both resting and activated cell populations, in human and mouse cells. ADDITIONAL NOTES I have talked to tech support and found out the antibody was tested against transfected cells. I need to know if you have data demonstrating this antibody can recognize phisological levels of APRIL in either human or murine cells.

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Answer

Thank you for your enquiry and patience. Regarding ab16085 (APRIL antibody), this antibody has not been tested on endogeneous APRIL. This would be difficult because APRIL is rapidly cleaved in the cells by furin and secreted. ab16085 was tested in FACS using HEK 293T cells that were transfected with an espression plasmid enabling surface expression of a non-cleavable mouse APRIL. I apologize that this information was missing from our datasheet - we will update this datasheet. Could you please let me know if you have tested this antibody in immunocytochemistry experiments? I am sorry that the product you received did not perform as stated on the datasheet. Since certain information was missing from the ab16085 datasheet, I am going to offer you a free of charge replacement order. Please let me know which Abcam product you would like to receive as a replacement. I apologize for the inconvenience and I look forward to your reply.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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