Overview

  • Product nameAnti-Semaphorin 7a antibody
    See all Semaphorin 7a primary antibodies
  • Description
    Rabbit polyclonal to Semaphorin 7a
  • Specificityab23578 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 applicationsSuitable for: WB, ICC/IF, IHC-FoFr, IHC-Fr, IHC-P, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat
    Predicted to work with: Cow
  • Immunogen

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

    (Peptide available as ab30844.)

  • Positive control
    • Developing neurons, differentiated from neural precursor cells cultured from embryonic day 13 mouse neocortex (neurons were cultured on poly-D-lysine in the presence of Neurobasal Medium containing B27 supplement)
  • General notesNegative control: HEK cells

Properties

Associated products

Applications

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.

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 Use a concentration of 1 - 25 µg/ml.
IHC-FoFr 1/100. (PMID 19741144)
IHC-Fr 1/100. (PMID 19741144)
IHC-P 1/100. (see Abreview)
IP Use at an assay dependent concentration.

Target

  • FunctionPlays 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 specificityDetected 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 similaritiesBelongs to the semaphorin family.
    Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
    Contains 1 PSI domain.
    Contains 1 Sema domain.
  • Cellular localizationCell membrane. Detected in a punctate pattern on the cell membrane of basal and supra-basal skin keratinocytes.
  • Information by UniProt
  • Database links
  • 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

Anti-Semaphorin 7a antibody images

  • 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).
  • 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).

  • 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
    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.

    See Abreview

  • 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.

References for Anti-Semaphorin 7a antibody (ab23578)

This product has been referenced in:
  • Chen Y  et al. The neural guidance receptor Plexin C1 delays melanoma progression. Oncogene 32:4941-9 (2013). Read more (PubMed: 23160370) »
  • Klisch TJ  et al. In vivo Atoh1 targetome reveals how a proneural transcription factor regulates cerebellar development. Proc Natl Acad Sci U S A 108:3288-93 (2011). ICC/IF ; Mouse . Read more (PubMed: 21300888) »

See all 7 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Ms. Nicole Gatzemeier

Verified customer

Submitted Feb 15 2011

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 ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
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%
Username

Dr. Randal Moldrich

Verified customer

Submitted Mar 21 2006

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