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Anti-Semaphorin 7a antibody (ab23578)

  • Datasheet
  • SDS
Reviews (2)Q&A (1)References (17)

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Western blot - Anti-Semaphorin 7a antibody (ab23578)
  • Immunocytochemistry/ Immunofluorescence - Anti-Semaphorin 7a antibody (ab23578)
  • Immunocytochemistry/ Immunofluorescence - Anti-Semaphorin 7a antibody (ab23578)
  • Immunoprecipitation - Anti-Semaphorin 7a antibody (ab23578)

Key features and details

  • Rabbit polyclonal to Semaphorin 7a
  • Suitable for: WB, ICC/IF, IP
  • Reacts with: Mouse, Rat
  • Isotype: IgG

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Overview

  • Product name

    Anti-Semaphorin 7a antibody
    See all Semaphorin 7a primary antibodies
  • Description

    Rabbit polyclonal to Semaphorin 7a
  • Host species

    Rabbit
  • Specificity

    ab23578 detects Semaphorin 7a protein at 75 kDa by WB. Semaphorin 7a is a protein that is tethered to the cell membrane via a GPI-anchor and is also glycosylated, therefore it is not surprising that in WB we see some additional bands at ~50 and 100 kDa which are likely to be cleavage fragments and glycosylated protein respectively. Preincubation with the immunizing peptide ab30844 is successful in blocking antibody staining.
  • Tested Applications & Species

    Application Species
    ICC/IF
    Mouse
    Rat
    IP
    Mouse
    WB
    Mouse
    See all applications and species data
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Mouse Semaphorin 7a.

    Read Abcam's proprietary immunogen policy (Peptide available as ab30844.)
  • General notes

    Negative control: HEK cells

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Blood
    • Other
    • Neuroscience
    • Neurology process
    • Growth and Development
    • Axonal Guidance Proteins
    • Neuroscience
    • Neurology process
    • Neurogenesis

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human Semaphorin 7a protein (Fc Chimera His Tag) (ab227401)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab23578 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Mouse
Rat
IP
Mouse
WB
Mouse
All applications
Cow
Application Abreviews Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 50,75,100 kDa (predicted molecular weight: 75 kDa).
ICC/IF (1)
Use a concentration of 1 - 25 µg/ml.
IP
Use at an assay dependent concentration.
Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 50,75,100 kDa (predicted molecular weight: 75 kDa).
ICC/IF
Use a concentration of 1 - 25 µg/ml.
IP
Use at an assay dependent concentration.

Target

  • Function

    Plays an important role in integrin-mediated signaling and functions both in regulating cell migration and immune responses. Promotes formation of focal adhesion complexes, activation of the protein kinase PTK2/FAK1 and subsequent phosphorylation of MAPK1 and MAPK3. Promotes production of proinflammatory cytokines by monocytes and macrophages. Plays an important role in modulating inflammation and T-cell-mediated immune responses. Promotes axon growth in the embryonic olfactory bulb. Promotes attachment, spreading and dendrite outgrowth in melanocytes.
  • Tissue specificity

    Detected in skin keratinocytes and on endothelial cells from skin blood vessels (at protein level). Expressed in fibroblasts, keratinocytes, melanocytes, placenta, testis, ovary, spleen, brain, spinal chord, lung, heart, adrenal gland, lymph nodes, thymus, intestine and kidney.
  • Sequence similarities

    Belongs to the semaphorin family.
    Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
    Contains 1 PSI domain.
    Contains 1 Sema domain.
  • Cellular localization

    Cell membrane. Detected in a punctate pattern on the cell membrane of basal and supra-basal skin keratinocytes.
  • Target information above from: UniProt accession O75326 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 20361 Mouse
    • Entrez Gene: 315711 Rat
    • SwissProt: Q9QUR8 Mouse
    • Unigene: 335187 Mouse
    • Alternative names

      • CD 108 antibody
      • CD108 antibody
      • CD108 antigen antibody
      • CDw108 antibody
      • H Sema K1 antibody
      • H Sema L antibody
      • H-SEMA-K1 antibody
      • H-Sema-L antibody
      • JMH antibody
      • JMH blood group antigen antibody
      • John Milton Hagen blood group H Sema K1 antibody
      • John Milton Hargen human blood group Ag antibody
      • John-Milton-Hargen human blood group Ag antibody
      • MGC126692 antibody
      • MGC126696 antibody
      • SEM7A_HUMAN antibody
      • Sema domain immunoglobulin domain (Ig) and GPI membrane anchor 7A antibody
      • Sema K1 antibody
      • Sema L antibody
      • SEMA7A antibody
      • SEMAK1 antibody
      • SEMAL antibody
      • Semaphorin 7A GPI membrane anchor antibody
      • Semaphorin K1 antibody
      • Semaphorin L antibody
      • Semaphorin-7A antibody
      • Semaphorin-K1 antibody
      • Semaphorin-L antibody
      • Semaphorin7a antibody
      see all

    Images

    • Western blot - Anti-Semaphorin 7a antibody (ab23578)
      Western blot - Anti-Semaphorin 7a antibody (ab23578)
      All lanes : Anti-Semaphorin 7a antibody (ab23578) at 1 µg/ml

      Lane 1 : Brain (Mouse) Tissue Lysate (ab27253)
      Lane 2 : Mouse brain tissue lysate - total protein (0 days) (ab7188)
      Lane 3 : Brain (Mouse) Tissue Lysate (ab27253) with Mouse Semaphorin 7a peptide (ab30844) at 1 µg/ml
      Lane 4 : Mouse brain tissue lysate - total protein (0 days) (ab7188) with Mouse Semaphorin 7a peptide (ab30844) at 1 µg/ml

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Rabbit IgG secondary antibody (ab28446) at 1/10000 dilution

      Performed under reducing conditions.

      Predicted band size: 75 kDa
      Observed band size: 75 kDa
      Additional bands at: 100 kDa (possible glycosylated form), 50 kDa (possible cleavage fragment)



      ab23578 detects Semaphorin 7a protein at 75kDa by WB. Semaphorin 7a is a protein that is tethered to the cell membrane via a GPI-anchor and is also glycosylated, therefore it is not surprising that we see some additional bands at ~50 and 100 kDa which are likely to be cleavage fragments and glycosylated protein respectively.

    • Immunocytochemistry/ Immunofluorescence - Anti-Semaphorin 7a antibody (ab23578)
      Immunocytochemistry/ Immunofluorescence - Anti-Semaphorin 7a antibody (ab23578)
      ICC/IF image of ab23578 stained PC12 cells. The cells were 4% formaldehyde fixed (10 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 (ab23578, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue).
    • Immunocytochemistry/ Immunofluorescence - Anti-Semaphorin 7a antibody (ab23578)
      Immunocytochemistry/ Immunofluorescence - Anti-Semaphorin 7a antibody (ab23578)This image is courtesy of Randal Moldrich, CNRS UMR7637, ESPCI, France

      ab23578 detecting Semaphorin 7a in developing mouse neurons, differentiated from neural precursor cells cultured from embryonic day 13 neocortex; neurons were cultured on poly-D-lysine in the presence of Neurobasal Medium containing B27 supplement. Immunocytochemistry: all steps were performed in PBS. Cultures were fixed in 4% PFA for 15min, permeabilised with 0.1% Triton X-100 for 10min and blocked with 5% BSA, 0.1% Triton X-100 for 45min. ab23578 was incubated at 5µg/ml (12h in 5% BSA, 0.1% Triton X-100) at 4°C. Goat anti-rabbit AlexaFluor 488 was used as secondary antibody at 1/400 for 1hr at RT (in 5% BSA, 0.1% Triton X-100). Treated cultures were mounted on glass coverslips with Mowiol. ab23578 immunostaining (green colour) was neuronal and for the most part at the membrane with some cytosolic staining observed. No nuclear staining was observed, confirmed with a nuclear counterstain (To-pro-3; data not shown).

    • Immunoprecipitation - Anti-Semaphorin 7a antibody (ab23578)
      Immunoprecipitation - Anti-Semaphorin 7a antibody (ab23578)
      Semaphorin 7a was immunoprecipitated using 0.5mg Mouse Brain whole tissue lysate, 5µg of Rabbit polyclonal to Semaphorin 7a and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
      The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
      Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab23578.
      Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
      Band: 75kDa: Semaphorin 7a.

    Protocols

    • Immunoprecipitation protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (17)

    Publishing research using ab23578? Please let us know so that we can cite the reference in this datasheet.

    ab23578 has been referenced in 17 publications.

    • Köhler D  et al. Red blood cell-derived semaphorin 7A promotes thrombo-inflammation in myocardial ischemia-reperfusion injury through platelet GPIb. Nat Commun 11:1315 (2020). PubMed: 32161256
    • Hu S  et al. Semaphorin 7A Promotes VEGFA/VEGFR2-Mediated Angiogenesis and Intraplaque Neovascularization in ApoE-/- Mice. Front Physiol 9:1718 (2018). PubMed: 30555351
    • Chen X  et al. Anti-Semaphorin-7A single chain antibody demonstrates beneficial effects on pulmonary inflammation during acute lung injury. Exp Ther Med 15:2356-2364 (2018). PubMed: 29456642
    • Chabrat A  et al. Transcriptional repression of Plxnc1 by Lmx1a and Lmx1b directs topographic dopaminergic circuit formation. Nat Commun 8:933 (2017). PubMed: 29038581
    • Jongbloets BC  et al. Stage-specific functions of Semaphorin7A during adult hippocampal neurogenesis rely on distinct receptors. Nat Commun 8:14666 (2017). WB ; Mouse . PubMed: 28281529
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-3 of 3 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Semaphorin 7a antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (spinal cord)
    Specification
    spinal cord
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citratbuffer pH6
    Permeabilization
    No
    Blocking step
    Endogenous peroxidase was inhibited by 1% H2O2in methanol for 15min. Sections were pretreated with 10% normal goat serum as blocking agent for 15 minute(s) · Concentration: 10% · Temperature: 22°C
    Read More

    Ms. Nicole Gatzemeier

    Verified customer

    Submitted Feb 15 2011

    Question

    BATCH NUMBER 170444 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Low signal. I wonder if we could get another lot for this antibody. We would appreciate if we could have small amount of antibody from different lot so that we could compare. SAMPLE Frozen section. PRIMARY ANTIBODY x100, x1000 DETECTION METHOD IF POSITIVE AND NEGATIVE CONTROLS USED No primary antibodies. We are using tissue that is known to express this antigen. FIXATION OF SAMPLE 4%PFA ANTIGEN RETRIEVAL Did not. Do we need to perform this for ab23578? There was no description in the data sheet for antigen retrieval. PERMEABILIZATION STEP 0.3% Triton X-100, 30 min BLOCKING CONDITIONS 2% donkey or goat serum SECONDARY ANTIBODY Alexa 594 or Alexa 488 against rabbit IgG HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? dilution

    Read More

    Abcam community

    Verified customer

    Asked on Apr 18 2006

    Answer

    Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. We recommend that this antibody is applied at a dilution range of 5 - 25ugml-1. This corresponds to a dilution range of 1:100 to 1:20. The low signal that you have been observing is possibly a consequence of the antibody being applied at a concentration that is too dilute. Furthermore the Abreview that has been left by one of our customers details an overnight incubation at 4oC. This is certainly likely to improve the signal that you have been obtaining. Antigen retrieval is not necessary for the application of immunohistochemistry to frozen sections. I hope this information helps. Please get back in touch with me should you continue to have difficulties with this antibody.

    Read More

    Abcam Scientific Support

    Answered on Apr 19 2006

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Semaphorin 7a antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (neurones differentiated from neural precursors)
    Specification
    neurones differentiated from neural precursors
    Fixative
    Paraformaldehyde
    Blocking step
    BSA as blocking agent for 40 minute(s) · Concentration: 5%
    Read More

    Dr. Randal Moldrich

    Verified customer

    Submitted Mar 21 2006

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

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