Overview

  • Product name
    Anti-Anterior Gradient 2 antibody
    See all Anterior Gradient 2 primary antibodies
  • Description
    Rabbit polyclonal to Anterior Gradient 2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Orangutan
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human Anterior Gradient 2.

    Read Abcam's proprietary immunogen policy (Peptide available as ab45078.)

  • Positive control
    • Recombinant Human Anterior Gradient 2 protein (ab64013) can be used as a positive control in WB. This antibody gave a positive signal in the following lysates: MCF7 whole cell lysate, Rat Small Intestine, Mouse Small Intestine

Properties

Applications

Our Abpromise guarantee covers the use of ab42765 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 20 kDa (predicted molecular weight: 20 kDa).
IHC-P Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • Function
    Required for MUC2 post-transcriptional synthesis and secretion. May play a role in the production of mucus by intestinal cells (By similarity). Proto-oncogene that may play a role in cell migration, cell differentiation and cell growth.
  • Tissue specificity
    Expressed strongly in trachea, lung, stomach, colon, prostate and small intestine. Expressed weakly in pituitary gland, salivary gland, mammary gland, bladder, appendix, ovary, fetal lung, uterus, pancreas, kidney, fetal kidney, testis, placenta, thyroid gland and in estrogen receptor (ER)-positive breast cancer cell lines.
  • Sequence similarities
    Belongs to the AGR family.
  • Cellular localization
    Secreted. Endoplasmic reticulum.
  • Information by UniProt
  • Database links
  • Alternative names
    • AG 2 antibody
    • AG 2 protein antibody
    • AG-2 antibody
    • AG2 antibody
    • AG2 protein antibody
    • AGR 2 antibody
    • AGR2 antibody
    • AGR2_HUMAN antibody
    • Anterior gradient 2 homolog (Xenopus laevis) antibody
    • anterior gradient 2 homolog antibody
    • Anterior gradient 2, protein disulphide isomerase family member antibody
    • Anterior gradient 2, Xenopus, homolog of antibody
    • Anterior gradient homolog 2 antibody
    • Anterior gradient protein 2 homolog antibody
    • Epididymis secretory protein Li 116 antibody
    • GOB 4 antibody
    • GOB4 antibody
    • HAG 2 antibody
    • hAG-2 antibody
    • hAG2 antibody
    • HAG2/R antibody
    • HEL S 116 antibody
    • HPC 8 antibody
    • hPC8 antibody
    • PDIA17 antibody
    • Protein disulfide isomerase family A member 17 antibody
    • secreted cement gland homolog antibody
    • Secreted cement gland protein XAG 2 homolog antibody
    • Secreted cement gland protein XAG-2 homolog antibody
    • Secreted cement gland protein XAG2 homolog antibody
    • XAG 2 antibody
    • XAG2 antibody
    see all

Images

  • All lanes : Anti-Anterior Gradient 2 antibody (ab42765) at 1 µg/ml

    Lane 1 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 2 : Small Intestine (Mouse) Tissue Lysate
    Lane 3 : Small Intestine (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 20 kDa
    Observed band size: 19 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 48 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 30 seconds
  • ICC/IF image of ab42765 stained HeLa cells. The cells were 4% PFA 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 (ab42765, 5µg/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) at a concentration of 1.43µM.

References

ab42765 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Question
Answer

Thank you for contacting us. Your credit note ID is XXXXXXX. I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways: (1) Redeemed against the original invoice if this hasn't already been paid. (2) Held on the account for use against a future order. (3) A full refund can be offered where no other invoices are outstanding. Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge. To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website. The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB. I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice  

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Answer

Thank you for providing further details. I am sorry to hear this antibody is not providing satisfactory results. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Having reviewed the details, I believe this product should have given satisfactory results. It appears that you may have received a faulty vial. I apologize for the inconvenience and am pleased to offer you a free of charge replacement, credit note, or refund in compensation. Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.  

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Answer

Thank you for your enquiry regarding ab42765 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. Additionally, thank you for supplying an image, as this has been very beneficial in understanding your concerns. The protocol looks fine to me however I would like to ask few questions which would help me to identify the source of the problem - What is the difference between the top 2 images and bottom 2 images? Are these using different dilutions or conc. of antibodies? In top image with ab42765 there is a faint band at the expected band size; this faint band can be further clarified using troubleshooting steps e.g using high conc. of primary antibody or protein loading. Could you specify if you have optimized any step in protocol? In the 2nd image the antibody is detecting the band at right size which is just on the red mark. Do you think this is not a right band? I am unfortunately unable to explain the identity of strong bands at 30 and 50 kDa; which presumably are the non specific bands. Could you specify which cell line lysates was used for whole cell lysate preparation? In order to proceed further in this case I will look forward to hearing from you soon.

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Application
Western blot
Sample
Mouse Cell lysate - whole cell (P388D1, macrophages)
Loading amount
10 µg
Specification
P388D1, macrophages
Gel Running Conditions
Reduced Denaturing (17%)
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Sep 10 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (epithelial colorectal adenocarcinoma cells (Caco))
Loading amount
20 µg
Specification
epithelial colorectal adenocarcinoma cells (Caco)
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jul 09 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Prostate)
Specification
Prostate
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako Target Retreival Solution
Permeabilization
No
Blocking step
5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C

Mr. Antibody Solutions

Verified customer

Submitted May 25 2010

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

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