Overview

  • Product name
  • Description
    Rabbit polyclonal to Slit2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Human
    Predicted to work with: Xenopus laevis
  • Immunogen

    Synthetic peptide:

    YFIPGTEDYRSKLSGDC

    conjugated to KLH, corresponding to amino acids 484-500 of Human SLIT2.

  • Positive control
    • Chicken spinal cord whole cell lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab7665 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
ICC/IF Use at an assay dependent concentration.
WB 1/500 - 1/2000. Detects a band of approximately 165 kDa (predicted molecular weight: 170 kDa). Incubate primary antibody blocking buffer (5% goat serum + 0.5% non-fat dry milk in PBS) other blocking buffers cause a poor signal.
IHC-Fr Use at an assay dependent concentration. PubMed: 20498081

Target

  • Function
    Thought to act as molecular guidance cue in cellular migration, and function appears to be mediated by interaction with roundabout homolog receptors. During neural development involved in axonal navigation at the ventral midline of the neural tube and projection of axons to different regions. SLIT1 and SLIT2 seem to be essential for midline guidance in the forebrain by acting as repulsive signal preventing inappropriate midline crossing by axons projecting from the olfactory bulb. In spinal chord development may play a role in guiding commissural axons once they reached the floor plate by modulating the response to netrin. In vitro, silences the attractive effect of NTN1 but not its growth-stimulatory effect and silencing requires the formation of a ROBO1-DCC complex. May be implicated in spinal chord midline post-crossing axon repulsion. In vitro, only commissural axons that crossed the midline responded to SLIT2. In the developing visual system appears to function as repellent for retinal ganglion axons by providing a repulsion that directs these axons along their appropriate paths prior to, and after passage through, the optic chiasm. In vitro, collapses and repels retinal ganglion cell growth cones. Seems to play a role in branching and arborization of CNS sensory axons, and in neuronal cell migration. In vitro, Slit homolog 2 protein N-product, but not Slit homolog 2 protein C-product, repels olfactory bulb (OB) but not dorsal root ganglia (DRG) axons, induces OB growth cones collapse and induces branching of DRG axons. Seems to be involved in regulating leukocyte migration.
  • Tissue specificity
    Fetal lung and kidney, and adult spinal cord. Weak expression in adult adrenal gland, thyroid, trachea and other tissues examined.
  • Sequence similarities
    Contains 1 CTCK (C-terminal cystine knot-like) domain.
    Contains 7 EGF-like domains.
    Contains 1 laminin G-like domain.
    Contains 20 LRR (leucine-rich) repeats.
    Contains 4 LRRCT domains.
    Contains 4 LRRNT domains.
  • Domain
    The leucine-rich repeat domain is sufficient for guiding both axon projection and neuronal migration, in vitro.
  • Cellular localization
    Secreted. The C-terminal cleavage protein is more diffusible than the larger N-terminal protein that is more tightly cell associated.
  • Information by UniProt
  • Database links
  • Alternative names
    • Drad 1 antibody
    • E030015M03Rik antibody
    • E130320P19Rik antibody
    • FLJ14420 antibody
    • OTTHUMP00000158695 antibody
    • OTTHUMP00000217852 antibody
    • OTTHUMP00000217853 antibody
    • OTTHUMP00000217854 antibody
    • Slil 3 antibody
    • Slil3 antibody
    • Slit 2 antibody
    • Slit homolog 2 (Drosophila) antibody
    • Slit homolog 2 antibody
    • Slit homolog 2 protein antibody
    • Slit homolog 2 protein C-product antibody
    • Slit-2 antibody
    • Slit2 antibody
    • SLIT2_HUMAN antibody
    see all

Images

  • Anti-Slit2 antibody (ab7665) at 1/1350 dilution + Chicken spinal cord whole cell lysate at 30 µg

    Secondary
    IRDye™800
    conjugated Goat anti-Rabbit IgG [H&L] at 1/10000 dilution

    Predicted band size: 170 kDa
    Observed band size: 165 kDa
    why is the actual band size different from the predicted?



    Molecular weight estimation was made by comparison to prestained molecular weight markers

  • ab7665 staining Slit2 in rat glioma cell line C6 by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed in paraformaldehyde, blocked with 0.5% BSA for 20 minutes at room temperature, then incubated with ab7665 at a 1/80 dilution for 16 hours at 4°C. The secondary used was a TRITC conjugated goat anti-rabbit polyclonal, used at a 1/400 dilution. Nuclei are counterstained with DAPI.

    See Abreview

  • ab7665 staining Slit2 in human glioblastoma cell line D54MG by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed in paraformaldehyde, permeabilized with 0.1% Triton X-100, blocked with 0.5% BSA for 20 minutes at room temperature, then incubated with ab7665 at a 1/80 dilution for 16 hours at 4°C. The secondary used was a TRITC conjugated goat anti-rabbit polyclonal, used at a 1/400 dilution. Nuclei are counterstained with DAPI.

    See Abreview

  • ab7665 staining Slit2 in human glioblastoma cell line by Immunocytochemistry/ Immunofluorescence.
    Cells were fixed in paraformaldehyde, permeabilized using 0,1% Triton X 100 in PBS, blocked with 0.5% BSA for 30 minutes at room temperature and then incubated with ab7665 at a 1/50 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-rabbit polyclonal used at a 1/400 dilution. Nuclei are counterstained with DAPI.

    See Abreview

References

This product has been referenced in:
  • Sakima M  et al. VEGFR-3 signaling is regulated by a G-protein activator, activator of G-protein signaling 8, in lymphatic endothelial cells. Exp Cell Res 368:13-23 (2018). Human . Read more (PubMed: 29649427) »
  • Piao JM  et al. MicroRNA-381 Favors Repair of Nerve Injury Through Regulation of the SDF-1/CXCR4 Signaling Pathway via LRRC4 in Acute Cerebral Ischemia after Cerebral Lymphatic Blockage. Cell Physiol Biochem 46:890-906 (2018). Read more (PubMed: 29669322) »
See all 9 Publications for this product

Customer reviews and Q&As

1-10 of 18 Abreviews or Q&A

Question
Answer

As discussed the mouse cross reactivity was added due to its citation in PMID 20498081. We unfortunately do not have any other mouse cross reactivity data available.

I have cheked the SLIT2 antibodies from different vendoes. The observed band was 130 kDa with mouse brain lysates.

Please be aware that SLIT2 protein is believed to form 2 cleavage products 130-140 kDa (N product) and 55 kDa (C product). (http://www.uniprot.org/uniprot/Q9R1B9). This antibody may show 130-140 kDa band with mouse lysates. You may need to check the literature about this.

Read More

Answer

Thank you very much for your email.

The preparation of tissue and cell lysates looks fine to me. The antibody should have worked, I however have following recommendations, I am sure these will help to improve the results.

- Heat lysates in sample buffer 5-10 minutes at 100C.
- ab7665 is a rabbit polyclonal antibody. Please use HRP conjugated ANTI- RABBIT secondary antibody.
- 1% NP-40 lysis buffer will give good results.

Please try these suggestions and let me know if the results do not improve.

Read More

Question
Answer

Thanks you very much for sending the details and image.

I have reviewed the protocol being used and also have checked the provide image; I am sorry I have few more questions to ask;

- How the cell lysates were prepared? What buffer was used? Was the buffer ice cold? For how long cells left in lysis buffer? have you centrifuged the lysates after preparation?
- How the GUT and Brain lysates were prepared?
- At what temperature the cell lysates were heated in sample buffer and for how long?
- For how long the membrane was blocked?
- Was the membrane (PVDF) activated with methanol/ Transfer buffer?
- What was the secondary antibody?

I have few suggestions for improvement:

- Please avoid touching the membrane with bare hands?
- Prepare the lysates in Ice cold RIPA buffer. The protocol we recommend is; after washing the cells in tissue culture plate with PBS, add lysis buffer (RIPA buffer), scrap of the cells and add into microcentrifuge tube, leave at 4C with constant agitation for 30 minutes, centrifuge at 16,000 g and discard the pellet.
- We recommend heating the lysates in sample buffer for 10 minutes at 100C.
- We recommend blocking the membrane for 1-2 hours in 5%BSA at RT.
- We recommend soaking the membrane for 5-10 minute in transfer buffer.
- We recomend preparing the lysates when cells 70-80% confluent.

Please follow these steps result will be much cleaner. If the result do not improve please do not heasitate to contact em again for further assistance.

Read More

Question
Answer

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.


I look forward to receiving your reply.

Read More

Answer

Thank you for contacting us.

The Anti-Slit2 antibody (ab7665) should detect a band at ˜165 kDa indeed, not 55 kDa.

In order to understand the detection of a band at 55 kDa, have you performed a "no primary" control (incubation with the secondary only)? The band at 55 kDa may be due to the secondary antibodyand not the primary.

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

Read More

Question
Answer

DISCOUNT CODE: XXXXXXXXXX
Expiration date: 2012-07-05

I am very pleased to hear you would like to accept our offer and test ab7665 in IHC-P. This code will give you 1 free [PRIMARY ANTIBODY] before the expiration date. To redeem this offer, please submit an Abreview for IHC-P and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: https://www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Question
Answer

Thank you for your enquiry.

Here is a link to more information regarding our Abreview system: https://www.abcam.com/abreviews

I hope this is helpful. Please contact me again if you have any further questions.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human glioblastoma cell line)
Specification
human glioblastoma cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - 0,1% Triton X 100 in PBS
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jan 24 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (glioblastoma cell line C6)
Specification
glioblastoma cell line C6
Fixative
Paraformaldehyde
Permeabilization
Yes - 0,1% Triton X 100 in PBS
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jan 24 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (human glioblastoma cell line D54MG)
Specification
human glioblastoma cell line D54MG
Fixative
Paraformaldehyde
Permeabilization
Yes - 0,1% Triton X 100
Blocking step
BSA as blocking agent for 20 minute(s) · Concentration: 0.5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Aug 30 2011

1-10 of 18 Abreviews or Q&A

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