Overview

  • Product name
    Anti-GM130 antibody - Drosophila Golgi/Cis-Golgi Marker
    See all GM130 primary antibodies
  • Description
    Rabbit polyclonal to GM130 - Drosophila Golgi/Cis-Golgi Marker
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Drosophila melanogaster
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 750 to the C-terminus of Fruit fly (Drosophila melanogaster) GM130.

    Read Abcam's proprietary immunogen policy (Peptide available as ab32335.)

  • Positive control
    • Drosophila S2 Cells

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
    Constituents: PBS, 1% BSA

    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

Applications

Our Abpromise guarantee covers the use of ab30637 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 a concentration of 1 µg/ml.
WB Use a concentration of 0.1 µg/ml. Detects a band of approximately 95 kDa (predicted molecular weight: 95 kDa).

Target

  • Function
    Golgi auto-antigen; probably involved in maintaining cis-Golgi structure.
  • Sequence similarities
    Belongs to the GOLGA2 family.
  • Domain
    Extended rod-like protein with coiled-coil domains.
  • Cellular localization
    Golgi apparatus > Golgi stack membrane.
  • Information by UniProt
  • Alternative names
    • 130 kDa cis Golgi matrix protein antibody
    • 130 kDa cis-Golgi matrix protein antibody
    • Cis golgi matrix protein GM130 antibody
    • GM130 antibody
    • Gm130 autoantigen antibody
    • GOGA2_HUMAN antibody
    • GOLGA 2 antibody
    • Golga2 antibody
    • Golgi autoantigen antibody
    • Golgi autoantigen golgin subfamily a 2 antibody
    • Golgi matrix protein GM130 antibody
    • Golgin 95 antibody
    • golgin A2 antibody
    • Golgin subfamily a 2 antibody
    • Golgin subfamily A member 2 antibody
    • Golgin-95 antibody
    • MGC20672 antibody
    • SY11 protein antibody
    see all

Images

  • The image shows staining of Drosophila S2 cells using ab30637 to cis-Golgi/Golgi marker, GM130. For comparison, S2 cells were stained with an antibody to the trans-Golgi marker, GCC88. Gm130 stained the cis-Golgi region and appears to be a superior Golgi marker to GCC88.
  • All lanes : Anti-GM130 antibody - Drosophila Golgi/Cis-Golgi Marker (ab30637) at 0.1 µg/ml

    Lane 1 : Drosophila melanogaster S2 cell extract
    Lane 2 : Drosophila melanogaster S2 cell extract + specific siRNA

    Predicted band size: 95 kDa
    Observed band size: 95 kDa

  • ab30637 at a 1/100 dilution staining Golgi/Cis-Golgi in Drosophila S2R+ cells by Immunocytochemistry/ Immunofluorescence.
    Expression of recombinant wild type pgant3 (pIB-pgant3-V5), pgant3m1 (pIB-m1-V5), and pgant3m2 (pIB-m2-V5) in Drosophila S2R+ cells. Cells were then stained for V5 and the Golgi marker GM130 revealing Golgi localization of wild type pgant3 and pgant3m1.
  • All lanes : Anti-GM130 antibody - Drosophila Golgi/Cis-Golgi Marker (ab30637) at 1 µg/ml

    Lane 1 : Schneider L2 whole cell lysate (ab14893)
    Lane 2 : Schneider L2 whole cell lysate (ab14893) with D. melanogaster GM130 peptide (ab32335) at 1 µg/ml

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Predicted band size: 95 kDa
    Observed band size: 105 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 70 kDa. We are unsure as to the identity of these extra bands.

References

This product has been referenced in:
  • Gerards M  et al. Intracellular vesicle trafficking plays an essential role in mitochondrial quality control. Mol Biol Cell N/A:N/A (2018). Read more (PubMed: 29343549) »
  • Gordon DE  et al. VAMP3/Syb and YKT6 are required for the fusion of constitutive secretory carriers with the plasma membrane. PLoS Genet 13:e1006698 (2017). Read more (PubMed: 28403141) »
See all 39 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (4-12%)
Sample
Zebrafish Tissue lysate - whole (total larvae, 4 days post fertilization)
Specification
total larvae, 4 days post fertilization
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Dr. Evgenia Salta

Verified customer

Submitted Oct 27 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Sample
Fruit fly (Drosophila melanogaster) Cell (Whole Ovary/egg chamber)
Specification
Whole Ovary/egg chamber
Permeabilization
Yes - 1% Triton-X100 2h 1% BSA
Fixative
Paraformaldehyde

Dr. Irina Catrina

Verified customer

Submitted Jun 06 2014

Question

here you are.



1) Abcam product code ab30637

2) Abcam order reference number or product batch number LOT GR65234-1

3) Description of the problem No specific staining

4) Sample preparation:

Species Dme Salivary Glands

Type of sample: PFA fixed salivary gland

Sample preparation PFA fix, PBS + Triton-X washes, PBS +Triton-X + BSA (BBT) block, then staining

Positive control older batch ab30637 antibody

Negative control none

5) Fixation step

Yes

If yes: Fixative agent and concentration PFA 4%

Fixation time 20’

Fixation temperature ambient

6) Antigen retrieval method

7) Permeabilization method: Triton-X

Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?

Permeabilizing agent and concentration: Triton-X 0.2%

8) Blocking agent (eg BSA, serum…): BSA

Concentration 0.1 %

Blocking time 45’

Blocking temperature ambient

9) Endogenous peroxidases blocked? no

Endogenous biotins blocked? no

10) Primary antibody (If more than one was used, describe in “additional notes”) :

Concentration or dilution 50 ng/ml or 10ng/ml

Diluent buffer BBT

Incubation time o/n 4C

11) Secondary antibody: 488 or 568 alexa

Species: goat or donkey

Reacts against: rabbit

Concentration or dilution 1/400

Diluent buffer BBT

Incubation time 2h

Fluorochrome or enzyme conjugate

12) Washing after primary and secondary antibodies:

Buffer BBT

Number of washes 4*30’ and 4*15’

13) Detection method confocal

14) How many times have you run this staining? 6x

Do you obtain the same results every time? yes

What steps have you altered to try and optimize the use of this antibody? Different []

Read More
Answer

Thank you for your enquiry regarding ab30637 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

After reading through the detailed protocol you kindly forwarded to Abcam, I would like to make the following comments/suggestions:

1) This is a polyclonal antibody and there may be batch-to-batch variations. I would suggest increasing the working concentration to 1 µg/ml (overnight incubation at 4oC) as suggested on the on-line datasheet under the Application note.

2) Does the detection system work fine? Have you used it successfully with another primary antibody? Have you run a no primary - only secondary antibody - control to see if any of the non-specific bands are due to the secondary or not?

I hope this will be useful for you. Should you still have any problem with this antibody after following these suggestions, then please do not hesitate to contact our Technical Department again.

Read More

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

Read More

Answer

Thank you for contacting Abcam.

I would like to thank you for your response. If the product has gone off we will gladly replace it. However the procedure for this is that we have the protocol used and troubleshooting steps that you have performed for our records. I do apologize for any inconveniences this might cause. If you would you be able to send that information to me I would be able to being processing this for you.

Please let me know if you have any questions or concerns.

Read More

Answer

Thank you for contacting us.

I am sorry to hear of the difficulties that you have been experiencing using this product. Would you mind sending me the details of your protocol including the tissue type you are working with and your sample preparations? Could you also include any antigen retrieval steps you use, blocking solution and incubation information and the antibody concentrations that you have tried. Any images that you may have and any steps that you have changed while troubleshooting would be extremely helpful as well.

It seems unlikely that your protocol is the issue here but these details will not only help me to better understand your difficulties, they are also used for internal testing of the products if deemed necessary. If we are unable to solve the problems you are experiencing we will replace or refund the antibody according to our Abpromise guarantee.


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

Answer

Thanks you very much for sending us more details about the experiment. I am sorry I am not sure why the antibody staining appears spotty. This antibody was used in many publications without problem. So perhaps you have received a bad vial of this product or somehow the sub cellular architecture of the Golgi apparatus collapsed and formed blubs that now appears as spots. To further rule out the problem because of bad vial, I can either send you a new free of charge vial of ab30637 from different lot or a new antibody against the same target. Could you specify which option you would like to go for? I will look forward to hearing from you soon.  

Read More

Question
Answer

Thank you for contacting us and for sending the images. I believe the problem is not due to antibody so I would suggest the following - The antibody is showing consistent results in cells outside the marked area. It looks similar as publication and image on the datasheet. So I am presuming that antibody is fine. However I do not know the reason of blobs that appear in cell centre. These could be due to different reason so may be protocol optimizations can help. My recommendations are - Use higher dilution of primary and secondary antibody. - Increase blocking time from 30 minutes to 1 -1.5 hours. - Could I ask were the cells healthy? - Could you provide details of fixative agent used; how the fixation was performed? - Could you send us an image of no DAPI control image? I look forward to hearing from you soon.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Fruit fly (Drosophila melanogaster) Cell (eye disc)
Specification
eye disc
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% triton
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Apr 28 2008

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