Overview

  • Product name

  • Description

    Rabbit polyclonal to GLG1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, IHC-FoFr, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Chicken, Human
  • Immunogen

    Synthetic peptide corresponding to Human GLG1 aa 1150-1179 (C terminal) conjugated to keyhole limpet haemocyanin.

  • Positive control

    • MCF7 cell line and Mouse cerebellum tissue lysates; skeletal muscle tissue. This antibody gave a positive result when used in the following formaldehyde fixed cell line: HepG2.

Properties

Applications

Our Abpromise guarantee covers the use of ab103439 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 1/100 - 1/500. Predicted molecular weight: 135 kDa.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IHC-FoFr Use at an assay dependent concentration.
ICC/IF 1/200.

Images

  • ICC/IF image of ab103439 stained HepG2 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 ab103439 at 1/200 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 Goat anti-Rabbit IgG (H+L) (ab96899) used at a 1/250 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) at a concentration of 1.43µM.

  • Anti-GLG1 antibody (ab103439) at 1/100 dilution + MCF7 cell line lysates at 35 µg

    Predicted band size: 135 kDa

  • Anti-GLG1 antibody (ab103439) at 1/100 dilution + Mouse cerebellum tissue lysates at 35 µg

    Predicted band size: 135 kDa

  • ab103439 at 1/50 dilution, staining Golgi Complex in Human skeletal muscle by Immunohistochemistry, Formalin-fixed, Paraffin-embedded tissue, followed by peroxidase-conjugated secondary antibody and DAB staining.
  • ab103439 staining Golgi Complex in murine spinal cord motor neurons by Immunohistochemistry (PFA perfusion fixed frozen sections).
    Tissue was fixed in paraformaldehyde, permeabilized using 0.2% Triton/PBS, blocked with 2.5% serum for 1 hour at 20°C and then incubated with ab103439 at a 1/500 dilution for 12 hours at 4°C. The secondary used was an Alexa-Fluor 488 conjugated goat anti-rabbit polyclonal used at a 1/500 dilution.

    See Abreview

References

This product has been referenced in:

  • Crociara P  et al. Motor neuron degeneration, severe myopathy and TDP-43 increase in a transgenic pig model of SOD1-linked familiar ALS. Neurobiol Dis 124:263-275 (2018). Read more (PubMed: 30471417) »
  • Rostami J  et al. Human Astrocytes Transfer Aggregated Alpha-Synuclein via Tunneling Nanotubes. J Neurosci 37:11835-11853 (2017). Read more (PubMed: 29089438) »
See all 4 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Sample
Chicken Cell (Primary oculomotor culture)
Specification
Primary oculomotor culture
Permeabilization
Yes - PBS + 0.1% Tween20
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Oct 27 2014

Application
Western blot
Loading amount
100000 cells
Gel Running Conditions
Reduced Denaturing (10%)
Sample
Human Cell lysate - whole cell (Breast Cancer MDA-MB-231 Cells/ Melanoma MDA-MB-43)
Specification
Breast Cancer MDA-MB-231 Cells/ Melanoma MDA-MB-43
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 50% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 28 2014

Answer

Thank you for taking time to contact us and forward an image of your staining. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

I'm copying the lab's IHC-P protocol below. Could you take a look and see if it is similar to the protocol you've tested? If so, and you don't think there are any changes to make, I would be happy to offer you a replacement or refund.

Preparing Tissue Sections for Immunostaining:

 Fix the tissue in 10% formalin at 4°C overnight.

 Embed fixed tissue in paraffin.

 Mount tissue sections on slides.

 Clear the paraffin with xylene for ten minutes; move slides to a fresh dish of xylene for an additional ten minutes. NOTE: Perform all xylene washes in a fume hood!

 Rinse the slides twice for 2 minutes in 100% alcohols (18:1:1 100% ethanol: 100% methanol: 100% isopropanol).

 Rinse the slides twice for 2 minutes in a 95% solution of the 100% alcohols.

 Place slides in an 80% solution of the 100% alcohols for 2 minutes, followed by deionized water for 5 minutes.

 Rinse slides several times with fresh deionized water followed by another five minutes wash using fresh water.

Sodium Citrate Antigen Retrieval:

 Place slides in a glass slide holder and fill in the rest of the rack with blank slides (10 totals) to ensure even heating.

 Place rack in 600 ml of 10 mM sodium citrate, pH 6.0 in a glass 2 L-beaker. Mark a line at the top of the liquid on the beaker.

 Microwave for 20 min total, replacing evaporated water every 5 min.

 Cool slides for 20 min.

 Wash 4 × 3 min in ddH2O, and 3 min in 1 × PBS.

Blocking

 Block endogenous peroxidases by soaking slides in a solution of 90% methanol/3% H2O2 for 15 minutes at room temperature. Wash 3 × in PBS.

 Immerse slides in a dish containing blocking buffer (serum from host species of secondary antibody to be used, diluted 1:10 in TBS). Incubate at 37°C for one hour.

Incubation with Primary Antibodies

 Cover the tissue sections with primary antibody diluted in blocking buffer. Antibody is diluted 1:50 and 1:100. Incubate for 1 hour at 37°C.

 Blot excess liquid from slides and rinse three times in PBS for five minutes each wash.

Incubation with Secondary Antibodies

 Cover the tissue sections with secondary antibody diluted in blocking buffer according to manufacturer’s instructions. We routinely use prediluted universal secondary antibody (Jackson ImmunoResearch Laboratories). Incubate at 37°C for 30 min.

 Blot excess liquid and rinse twice in TBS for five minutes each wash.

Counterstaining and Visualization

 Counterstain with Hematoxylin.

 Rinse several times in deionized water. Blot excess water around tissue, then apply one drop of mounting media to tissue and place coverslip over slide. Seal with nail polish.

Citrate Solutions:

Description: Formalin or other aldehyde fixation forms protein cross-links that mask the antigenic sites in tissue specimens, thereby giving weak or false negative staining for immunohistochemical detection of certain proteins. The citrate based solution is designed to break the protein cross-links, thereby unmasking the antigens and epitopes in formalin-fixed and paraffin embedded tissue sections and enhancing the staining intensity of antibodies.

Sodium Citrate Buffer (10mM Sodium Citrate, 0.05% Tween 20, pH 6.0):

Tri-sodium citrate (dihydrate) --------- 2.94 g

Distilled water --------------------------- 1000 ml

Mix to dissolve. Adjust pH to 6.0 with 1N HCl and then add 0.5 ml of Tween 20 and mix well. Store this solution at room temperature for 3 months or at 4°C for longer storage.

Citrate Buffer (10mM Citric Acid, 0.05% Tween 20, pH 6.0):

Citric acid (anhydrous) ---------------- 1.92 g

Distilled water -------------------------- 1000 ml

Mix to dissolve. Adjust pH to 6.0 with 1N NaOH and then add 0.5 ml of Tween 20 and mix well. Store this solution at room temperature for 3 months or at 4°C for longer storage.

Washing Buffer:

1 × PBS:

NaCl --------------------------------------- 8 g

KCl ----------------------------------------- 0.2 g

Na2HPO4---------------------------------- 1.44 g

KH2PO4------------------------------------ 0.24 g

Distilled Water--------------------------- 800 mL

Adjust pH to 7.2 with HCl.

Adjust volume to 1 L with additional H2O

Normal Serum Blocking Buffer:

2% serum from host species of secondary antibody (blocking)

1% BSA (stabilizer)

0.1% cold fish skin gelatin (blocking)

0.1% Triton X-100 (penetration enhancer)

0.05% Tween 20 (detergent and surface tension reducer)

0.05% sodium azide (preservative)

Dissolve in 1 × PBS

Mix well and store at 4°C.

Avidin/Biotin Block:

Avidin 0.001% in 1 × PBS

Biotin 0.001% in 1 × PBS

Store these blocking solution at 4°C.

Primary Antibody Dilution Buffer:

1%BSA (stabilizer and blocking)

0.1% cold fish skin gelatin (blocking)

0.05% sodium azide (preservative)

0.01M PBS pH7.2

Peroxidase Blocking Solution (3% H2O2 in PBS):

30% H2O2 -------------------------- 2 ml

1 × PBS ----------------------------- 18 ml

Mix well and store at 4°C for up to 3 months.

This solution is recommended for paraffin sections

Should the suggestions not improve the results, please do let me know.

I hope this information is helpful, and I thank you for your cooperation.

Read More

Answer

Merci de nous avoir contactés.

Nous avons maintenantvalidé la commande. Le produit doit être envoyé de notre stock des Etats-Unis, vous devriez le recevoir mercredi 29 février.

Merci encore pour votre Abreview.

Read More
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Spinal cord motor neuron)
Specification
Spinal cord motor neuron
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
Yes - PBS Triton 0.1%
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 20°C

Dr. Georg Haase

Verified customer

Submitted Feb 24 2012

Question
Answer

Code de remise : ********
Date d’expiration : JJ MM AAAA

Je suis ravi que vous souhaitiez participerà notre offre et tester ab103439 en cytométrie en flux et/ou IHC-Fr. Le code de réduction correspondant est *********. Ce code de réduction est utilisable sous 120 jours et pour l’achat d’un autre anticorps primaire.

Vous devez dans un premier temps soumettre une Abreview avec vos résultats en flux et/ou IHC-Fr en notant ce code dans la section « Additional notes ». Ce code sera ainsi activé et vous pourrez par la suite contacter notre service clientèle par téléphone pour passer votre prochaine commande, en vous munissant des informations suivantes : code de réduction, numéro de commande de ab103439.

Nous restons à votre disposition pour plus d’informations concernant cette offre promotionnelle. Dans l’attente de recevoir votre Abreview nous vous souhaitons bonne chance dans vos recherches.

Termes et Conditions : www.abcam.com/collaborationdiscount.

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