58K Golgi Protein Antibody (58K-9) - BSA Free

Novus Biologicals | Catalog # NB600-412

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Bovine, Canine, Chicken

Cited:

Human, Mouse, Rat, Avian - Chicken

Predicted:

Chinese Hamster (92%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 58K-9

Format

BSA Free
View all 58K Golgi Protein Primary Antibodies »

Product Specifications

Immunogen

58K Golgi Protein purified from rat liver [UniProt# O88618]

Localization

Cytoplasmic and Golgi Apparatus

Marker

Golgi Apparatus Marker

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Theoretical MW

58 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for 58K Golgi Protein Antibody (58K-9) - BSA Free

Western Blot: 58K Golgi Protein Antibody (58K-9) [NB600-412]

Western Blot: 58K Golgi Protein Antibody (58K-9) [NB600-412]

Western Blot: 58K Golgi Protein Antibody (58K-9) [NB600-412] - Analysis of 58K golgi protein expression in rat liver tissue using NB600-412.
Immunocytochemistry/ Immunofluorescence: 58K Golgi Protein Antibody (58K-9) [NB600-412]

Immunocytochemistry/ Immunofluorescence: 58K Golgi Protein Antibody (58K-9) [NB600-412]

Immunocytochemistry/Immunofluorescence: 58K Golgi Protein Antibody (58K-9) [NB600-412] - HeLa cells, cultured on cover slips, were fixed with 10% formalin for 10 minutes and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were then incubated with 1:200 dilution of anti-58K Golgi Protein antibody (clone 58K-9) for overnight at 4C and detected with an anti-mouse Dylight 488 (Green) secondary at a 1:500 dilution. Alpha tubulin (DM1A) [NB100-690] was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at 1:500 dilution. Nuclei were counterstained with DAPI solution (Blue) [NBP2-31156]. Cells were imaged using a 40X objective. Antibody clone 58K-9 generated a specific signal in the Golgi apparatus of the cells.
Immunohistochemistry: 58K Golgi Protein Antibody (58K-9) [NB600-412]

Immunohistochemistry: 58K Golgi Protein Antibody (58K-9) [NB600-412]

58K-Golgi-Protein-Antibody-58K-9-Immunohistochemistry-NB600-412-img0011.jpg
Immunohistochemistry: 58K Golgi Protein Antibody (58K-9) [NB600-412]

Immunohistochemistry: 58K Golgi Protein Antibody (58K-9) [NB600-412]

Immunohistochemistry: 58K Golgi Protein Antibody (58K-9) [NB600-412] - Analysis of 58K Golgi Protein in mouse kidney using DAB with hematoxylin counterstain.
58K Golgi Protein Antibody (58K-9)

Immunocytochemistry/Immunofluorescence: 58K Golgi Protein Antibody (58K-9) [NB600-412] -

Immunocytochemistry/Immunofluorescence: 58K Golgi Protein Antibody (58K-9) [NB600-412] - Assessing Golgi morphology observed apoptosis in mBECs using Mouse 58K Golgi Protein at a dilution of 1:100 overnight at 4 °C. This image was taken on a Zeiss LSM 710 Confocal Microscope (META) with Zeiss Plan Apo 63x/1.40 oil. Image from verified customer review.
58K Golgi Protein Antibody (58K-9)

Immunocytochemistry/ Immunofluorescence: 58K Golgi Protein Antibody (58K-9) [NB600-412] -

Immunocytochemistry/ Immunofluorescence: 58K Golgi Protein Antibody (58K-9) [NB600-412] - Co-localization of Neu5Gc staining with markers for endosomes & Golgi in BMD & DMD muscle.(A) BMD muscle co-stained for Neu5Gc (green) & clathrin (red), a marker of endosomes. Merged image on right shows overlap of Neu5Gc & clathrin expression in yellow. Arrow marks several examples of co-staining. (B) DMD muscle co-stained for Neu5Gc (green) with LAMP1, a lysosomal marker, 58K Golgi, a Golgi marker, or calnexin, & endoplasmic reticulum marker, all in red, & DAPI (blue). Arrow marks region of coincident staining (yellow) for Neu5Gc & 58K Golgi. Bar is 50 µm for all panels in A & B. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/24505439), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for 58K Golgi Protein Antibody (58K-9) - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:50 - 1:250

Immunohistochemistry

1:100 - 1:200

Immunohistochemistry-Frozen

reported in scientific literature (PMID 24505439)

Immunohistochemistry-Paraffin

1:100 - 1:200

Western Blot

1:2000 - 1:5000
Application Notes
In Western blot a band is observed at ~58 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended. The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Reviewed Applications

Read 3 reviews rated 3.3 using NB600-412 in the following applications:

Formulation, Preparation, and Storage

Purification

Unpurified

Formulation

Ascites

Format

BSA Free

Preservative

0.1% Sodium Azide

Concentration

This product is unpurified. The exact concentration of antibody is not quantifiable.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: 58K Golgi Protein

The 58K Golgi protein is one of the most commonly used Golgi markers and has been proposed to implicate in linking Golgi to microtubules and it peripherally associates with cytosolic surface of Golgi membranes. This protein shows extensive overall sequence identity with porcine FTCD and has been suggested to represent rat FTCD (formiminotransferase-cyclodeaminase), a metabolic enzyme involved in the conversion of histidine to glutamic acid. FTCD's formiminotransferase activity facilitate transfer of a a formimino group from N-formimino-L-glutamic acid to tetrahydrofolate to generate glutamic acid and 5-formiminotetrahydrofolate, and the cyclodeaminase activity then converts the 5-formiminotetrahydrofolate to 5,10-methenyl tetrahydrofolate and ammonia. FTCD appears to be a dynamic component of Golgi and cycles between Golgi and earlier compartments of the secretory pathway. It also binds and promotes bundling of vimentin filaments originating from the Golgi, and mutations in this gene encoding for 58K Golgi Protein i.e. FTCD are associated with glutamate formiminotransferase deficiency (FIGLU-URIA).

Alternate Names

formimidoyltransferase cyclodeaminase, formimidoyltransferase-cyclodeaminase, formiminotransferase cyclodeaminase, Formiminotransferase-cyclodeaminase, human formiminotransferase cyclodeaminase, EC 4.3.1.410formiminotransferase-cyclodeaminase, LCHC1

Entrez Gene IDs

10841 (Human); 14317 (Mouse); 89833 (Rat)

Gene Symbol

FTCD

UniProt

O95954 (Human), Q91XD4 (Mouse), O88618 (Rat)

Additional 58K Golgi Protein Products

Product Documents for 58K Golgi Protein Antibody (58K-9) - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for 58K Golgi Protein Antibody (58K-9) - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for 58K Golgi Protein Antibody (58K-9) - BSA Free

Customer Reviews for 58K Golgi Protein Antibody (58K-9) - BSA Free (3)

3.3 out of 5
3 Customer Ratings
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Showing  1 - 3 of 3 reviews Showing All
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  • 58K Golgi Protein Antibody (58K-9)
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: Mouse Brain Endothelial Cells
    Species: Mouse
    Verified Customer | Posted 03/22/2023
    Primary mouse brain endothelial cells stained with 58K antibody (1:100). Staining is perinuclear but Golgi cannot be distinguished.
    58K Golgi Protein Antibody (58K-9) - BSA Free NB600-412
    Bio-Techne Response
    Thank you for reviewing our product. We are sorry to hear that this product did not perform as expected. We have been in touch with the customer to resolve this issue according to our Product Guarantee and to the customer’s satisfaction.
  • 58K Golgi Protein Antibody (58K-9)
    Name: Ali Almousawi
    Application: Immunocytochemistry
    Sample Tested: Cultured Mouse brain endothelial cells, Cultured Brain Endothelial cells and Mouse Brain Endothelial cells (mBECs)
    Species: Mouse
    Verified Customer | Posted 03/13/2023
    Assessing Golgi morphology observed apoptosis in mBECs using Mouse 58K Golgi Protein at a dilution of 1:100 overnight at 4 °C. This image was taken on a Zeiss LSM 710 Confocal Microscope (META) with Zeiss Plan Apo 63x/1.40 oil
    58K Golgi Protein Antibody (58K-9) - BSA Free NB600-412
  • 58K Golgi Protein Antibody (58K-9)
    Name: Anonymous
    Application: Immunocytochemistry
    Sample Tested: mouse vascular smooth muscle cell
    Species: Mouse
    Verified Customer | Posted 11/19/2018
    58K Golgi Protein Antibody (58K-9) - BSA Free NB600-412

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Protocols

View specific protocols for 58K Golgi Protein Antibody (58K-9) - BSA Free (NB600-412):


Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 30 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

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