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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)

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Western blot - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
  • Western blot - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
  • Flow Cytometry - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
  • Flow Cytometry - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
  • Immunoprecipitation - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
  • Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR27110-42] to CD98 - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

HRP HRP

Overview

  • Product name

    Anti-CD98 antibody [EPR27110-42] - BSA and Azide free
    See all CD98 primary antibodies

  • Description

    Rabbit monoclonal [EPR27110-42] to CD98 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type HeLa, HeLa whole cell lysate, K562, SW620, SH-SY5Y, Human kidney, Human breast, and Human skeletal muscle lysates IHC-P: Human kidney, Human colon carcinoma and Human pancreas tissues. ICC/IF: SLC3A2 and HeLa cells Flow Cyt: Wild-type HeLa and SH-SY5Y cells. IP: HeLa cell.

  • General notes

    ab307588 is the carrier free version of ab307587

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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 ab307588 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
Use at an assay dependent concentration. Predicted molecular weight: 70 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.
Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 70 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.

Target

  • Function

    Required for the function of light chain amino-acid transporters. Involved in sodium-independent, high-affinity transport of large neutral amino acids such as phenylalanine, tyrosine, leucine, arginine and tryptophan. Involved in guiding and targeting of LAT1 and LAT2 to the plasma membrane. When associated with SLC7A6 or SLC7A7 acts as an arginine/glutamine exchanger, following an antiport mechanism for amino acid transport, influencing arginine release in exchange for extracellular amino acids. Plays a role in nitric oxide synthesis in human umbilical vein endothelial cells (HUVECs) via transport of L-arginine. Required for normal and neoplastic cell growth. When associated with SLC7A5/LAT1, is also involved in the transport of L-DOPA across the blood-brain barrier, and that of thyroid hormones triiodothyronine (T3) and thyroxine (T4) across the cell membrane in tissues such as placenta. Involved in the uptake of methylmercury (MeHg) when administered as the L-cysteine or D,L-homocysteine complexes, and hence plays a role in metal ion homeostasis and toxicity. When associated with SLC7A5 or SLC7A8, involved in the cellular activity of small molecular weight nitrosothiols, via the stereoselective transport of L-nitrosocysteine (L-CNSO) across the transmembrane. Together with ICAM1, regulates the transport activity LAT2 in polarized intestinal cells, by generating and delivering intracellular signals. When associated with SLC7A5, plays an important role in transporting L-leucine from the circulating blood to the retina across the inner blood-retinal barrier.

  • Tissue specificity

    Expressed ubiquitously in all tissues tested with highest levels detected in kidney, placenta and testis and weakest level in thymus. During gestation, expression in the placenta was significantly stronger at full-term than at the mid-trimester stage. Expressed in HUVECS and at low levels in resting peripheral blood T-lymphocytes and quiescent fibroblasts. Also expressed in fetal liver and in the astrocytic process of primary astrocytic gliomas. Expressed in retinal endothelial cells and in the intestinal epithelial cell line C2BBe1.

  • Sequence similarities

    Belongs to the SLC3A transporter family.

  • Post-translational
    modifications

    Phosphorylation on Ser-406; Ser-408 or Ser-410 and on Ser-527 or Ser-531 by ecto-protein kinases favors heterotypic cell-cell interactions.

  • Cellular localization

    Apical cell membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. Localized to the plasma membrane when associated with SLC7A5 or SLC7A8. Localized to the placental apical membrane. Located selectively at cell-cell adhesion sites (By similarity). Colocalized with SLC7A8/LAT2 at the basolateral membrane of kidney proximal tubules and small intestine epithelia. Expressed in both luminal and abluminal membranes of brain capillary endothelial cells (By similarity).
  • Information by UniProt

  • Database links

    • Form

      There are 4 isoforms produced by alternative splicing.

    • Alternative names

      • 4F2 antibody
      • 4F2 cell surface antigen heavy chain antibody
      • 4F2 cell-surface antigen heavy chain antibody
      • 4F2 heavy chain antibody
      • 4F2 heavy chain antigen antibody
      • 4F2_HUMAN antibody
      • 4F2hc antibody
      • 4T2HC antibody
      • Antigen defined by monoclonal antibody 4F2 heavy chain antibody
      • Antigen identified by monoclonal antibodies 4F2 TRA1.10 TROP4 and T43 antibody
      • CD 98 antibody
      • CD98 antibody
      • CD98 antigen antibody
      • CD98 heavy chain antibody
      • CD98HC antibody
      • Heavy chain antibody
      • Lymphocyte activation antigen 4F2 large subunit antibody
      • MDU 1 antibody
      • MDU1 antibody
      • Monoclonal antibody 44D7 antibody
      • NACAE antibody
      • Slc3a2 antibody
      • Solute carrier family 3 (activators of dibasic and neutral amino acid transport) member 2 antibody
      • Solute carrier family 3 member 2 antibody
      see all

    Images

    • Western blot - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
      Western blot - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
      All lanes : Anti-CD98 antibody [EPR27110-42] (ab307587) at 1/1000 dilution

      Lane 1 : Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : SLC3A2 (CD98) knockout HeLa whole cell lysate
      Lane 3 : HeLa whole cell lysate
      Lane 4 : K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
      Lane 5 : SW620 (human colorectal adenocarcinoma epithelial cell) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

      Predicted band size: 70 kDa
      Observed band size: 70-130 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer and concentration: 5% NFDM/TBSTLow expression: SH-SY5Y

       Exposure time: 3 seconds

    • Western blot - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
      Western blot - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
      All lanes : Anti-CD98 antibody [EPR27110-42] (ab307587) at 1/1000 dilution

      Lane 1 : Human kidney tissue lysate
      Lane 2 : Human breast tissue lysate
      Lane 3 : Human skeletal muscle tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

      Predicted band size: 70 kDa
      Observed band size: 70-130 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer and concentration: 5% NFDM/TBST

      Low expression: skeletal muscle (PMID: 10574970, PMID: 11557028).

       

      The lanes 2 and 3 were developed using a high sensitivity ECL substrate.

       Exposure time: Lane 1: 6 seconds, lanes 2 and 3: 48 seconds.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)

      This data was developed using ab307587, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling CD98 with ab307587 at 1/5000 (0.099 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution. Positive staining on human kidney. The section was incubated with ab307587 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution.

      Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 10 mins

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)

      This data was developed using ab307587, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Human colon carcinom tissue labeling CD98 with ab307587 at 1/5000 (0.099 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution. Positive staining on human colon carcinoma (PMID: 29860836). The section was incubated with ab307587 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument.Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution.

      Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 10 mins

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)

      This data was developed using ab307587, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling CD98 with ab307587 at 1/5000 (0.099 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution. Positive staining on islet of human pancreas. The section was incubated with ab307587 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument.Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) at Ready to use dilution.

      Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 10 mins

    • Immunocytochemistry/ Immunofluorescence - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
      Immunocytochemistry/ Immunofluorescence - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)

      This data was developed using ab307587, the same antibody clone in a different buffer formulation.

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SLC3A2 (CD98) KO HeLa (SLC3A2 (CD98) knockout human cervix adenocarcinoma epithelial cell) (ab265708) cells labelling CD98 with ab307587 at 1/100 (4.97 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/mL) dilution (Green). Confocal image showing membrane and weak cytoplasmic staining in parental HeLa cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

    • Immunocytochemistry/ Immunofluorescence - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
      Immunocytochemistry/ Immunofluorescence - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)

      This data was developed using ab307587, the same antibody clone in a different buffer formulation.

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling CD98 with ab307587 at 1/100 (4.97 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/mL) dilution (Green). Confocal image showing membrane and weak cytoplasmic staining in HeLa cell line. Low expression: SH-SY5YImage was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution.

    • Flow Cytometry - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
      Flow Cytometry - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)

      This data was developed using ab307587, the same antibody clone in a different buffer formulation.

      Flow cytometric analysis of Wild-type HeLa (human cervix adenocarcinoma epithelial cell, Right) / SLC3A2 (CD98) knockout HeLa (Left) cells labelling CD98 with ab307587 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.Positive staining on HeLa cells (ab255928), while no staining on CD98 knockout HeLa cells (ab265708).

    • Flow Cytometry - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
      Flow Cytometry - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)

      This data was developed using ab307587, the same antibody clone in a different buffer formulation.

      Flow cytometric analysis of SH-SY5Y (human neuroblastoma epithelial cell, Left) / HeLa (human cervix adenocarcinoma epithelial cell, Right) cells labelling CD98 with ab307587 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.Low expression: SH-SY5Y.

    • Immunoprecipitation - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
      Immunoprecipitation - Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)

      This data was developed using ab307587, the same antibody clone in a different buffer formulation.

      CD98 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug with ab307587 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307587 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug

      Lane 2: abab307587 IP in HeLa whole cell lysate

      Lane 3:Rabbit monoclonal IgG (ab172730) instead of ab307587 in hela whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 1 second

    • Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)
      Anti-CD98 antibody [EPR27110-42] - BSA and Azide free (ab307588)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab307588? Please let us know so that we can cite the reference in this datasheet.

    ab307588 has not yet been referenced specifically in any publications.

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