Name: Anti-Lipocalin-2 / NGAL antibody [5G5]
SKU: ab23477
Rating: 4 (3 reviews)

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Western blot - Anti-Lipocalin-2 / NGAL antibody [5G5] (ab23477)

Key features and details

Mouse monoclonal [5G5] to Lipocalin-2 / NGAL
Suitable for: WB, IHC-FoFr, IHC-P
Knockout validated
Reacts with: Human
Isotype: IgG1

Product name

Anti-Lipocalin-2 / NGAL antibody [5G5]
See all Lipocalin-2 / NGAL primary antibodies
Description

Mouse monoclonal [5G5] to Lipocalin-2 / NGAL
Host species

Mouse
Specificity

ab23477 is specific for Lipocalin-2 / NGAL, native and denatured forms.
Tested applications

Suitable for: WB, IHC-FoFr, IHC-Pmore details
Species reactivity

Reacts with: Human
Immunogen

Full length native protein (purified) corresponding to Human Lipocalin-2/ NGAL. Full length native human neutrophil gelatinase associated lipocalin isolated from neutrophils.
Positive control
- WB: SW480 cell lysate. IHC-P: PaCa, chronic pancreatitis and normal pancreas tissue sections.
General notes

The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at 4°C or -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.40
Preservative: 0.097% Sodium azide
Constituents: 0.0268% PBS, 2.9% Sodium chloride
Concentration information loading...
Purity

Protein G purified
Clonality

Monoclonal
Clone number

5G5
Myeloma

x63-Ag8.653
Isotype

IgG1
Light chain type

kappa
Research areas

Compatible Secondaries
- Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
- Goat Anti-Mouse IgG H&L (HRP) (ab205719)
Isotype control
- Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
KO cell lines
- Human LCN2 knockout SW480 cell line (ab270486)
KO cell lysates
- Human LCN2 knockout SW480 cell lysate (ab270509)
Recombinant Protein
- Recombinant Mouse Lipocalin-2 / NGAL protein (ab95008)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab23477 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	(2)	1/1000. Predicted molecular weight: 22 kDa.
IHC-FoFr		Use at an assay dependent concentration.
IHC-P	(1)	Use at an assay dependent concentration.

Notes
WB 1/1000. Predicted molecular weight: 22 kDa.
IHC-FoFr Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.

Function

Iron-trafficking protein involved in multiple processes such as apoptosis, innate immunity and renal development. Binds iron through association with 2,5-dihydroxybenzoic acid (2,5-DHBA), a siderophore that shares structural similarities with bacterial enterobactin, and delivers or removes iron from the cell, depending on the context. Iron-bound form (holo-24p3) is internalized following binding to the SLC22A17 (24p3R) receptor, leading to release of iron and subsequent increase of intracellular iron concentration. In contrast, association of the iron-free form (apo-24p3) with the SLC22A17 (24p3R) receptor is followed by association with an intracellular siderophore, iron chelation and iron transfer to the extracellular medium, thereby reducing intracellular iron concentration. Involved in apoptosis due to interleukin-3 (IL3) deprivation: iron-loaded form increases intracellular iron concentration without promoting apoptosis, while iron-free form decreases intracellular iron levels, inducing expression of the proapoptotic protein BCL2L11/BIM, resulting in apoptosis. Involved in innate immunity, possibly by sequestrating iron, leading to limit bacterial growth.
Tissue specificity

Expressed in bone marrow and in tissues that are prone to exposure to microorganism. High expression is found in bone marrow as well as in uterus, prostate, salivary gland, stomach, appendix, colon, trachea and lung. Not found in the small intestine or peripheral blood leukocytes.
Sequence similarities

Belongs to the calycin superfamily. Lipocalin family.
Cellular localization

Secreted. Upon binding to the SLC22A17 (24p3R) receptor, it is internalized.
Target information above from: UniProt accession P80188 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3934 Human
- Omim: 600181 Human
- SwissProt: P80188 Human
- Unigene: 204238 Human
Alternative names
- 24p3 antibody
- 25 kDa alpha 2 microglobulin related subunit of MMP9 antibody
- 25 kDa alpha-2-microglobulin-related subunit of MMP-9 antibody
- Alpha 2 microglobulin related protein antibody
- HGNC:6526 antibody
- HNL antibody
- Lcn 2 antibody
- Lcn2 antibody
- Lipocalin-2 antibody
- Migration stimulating factor inhibitor antibody
- MSFI antibody
- Neutrophil gelatinase associated lipocalin antibody
- Neutrophil gelatinase associated lipocalin precursor antibody
- Neutrophil gelatinase-associated lipocalin antibody
- NGAL antibody
- NGAL_HUMAN antibody
- Oncogene 24p3 antibody
- Oncogenic lipocalin 24p3 antibody
- p25 antibody
- Siderocalin antibody
- siderocalin LCN2 antibody
- SV40 induced 24P3 protein antibody
- Uterocalin antibody
see all

Western blot - Anti-Lipocalin-2 / NGAL antibody [5G5] (ab23477)

All lanes : Anti-Lipocalin-2 / NGAL antibody [5G5] (ab23477) at 1/1000 dilution

Lane 1 : Wild-type SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate
Lane 2 : LCN2 knockout SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate
Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate
Lane 4 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 22 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab23477 observed at 23 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.

ab23477 was shown to react with Lipocalin-2 / NGAL in wild-type SW480 cells in Western blot with loss of signal observed in LCN2 knockout knockout cell line ab270486 (knockout cell lysate ab270509). Wild-type SW480 and LCN2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab23477 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [5G5] (ab23477)This image is courtesy of an anonymous Abreview

ab23477 staining human skin tissue sections by IHC-P. Sections were fixed in 10% neutral buffered formalin and subjected to heat mediated antigen retrieval prior to blocking with 1% BSA for 1 hour at 25°C. The primary antibody was diluted 1/50 and incubated with the sample for 20 hours at 4°C. A biotinylated horse anti-mouse antibody diluted 1/200 was used as the secondary.
See Abreview
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [5G5] (ab23477)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human PaCa tissue sections labelling Lipocalin 2 with ab23477.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [5G5] (ab23477)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human chronic pancreatits tissue sections labelling Lipocalin-2 / NGAL with ab23477.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [5G5] (ab23477)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human pancreas tissue sections labelling Lipocalin-2 / NGAL with ab23477.

Immunohistochemistry protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab23477? Please let us know so that we can cite the reference in this datasheet.

ab23477 has been referenced in 15 publications.

Cione E et al. Expression of MMP-2, MMP-9, and NGAL in Tissue and Serum of Patients with Vascular Aneurysms and Their Modulation by Statin Treatment: A Pilot Study. Biomolecules 10:N/A (2020). PubMed: 32111073
Guo Y et al. NGAL protects in nasopharyngeal carcinoma by inducing apoptosis and blocking epithelial-mesenchymal transition. Oncol Lett 19:3711-3718 (2020). PubMed: 32391093
Monisha J et al. NGAL is Downregulated in Oral Squamous Cell Carcinoma and Leads to Increased Survival, Proliferation, Migration and Chemoresistance. Cancers (Basel) 10:N/A (2018). PubMed: 29996471
Ku AT et al. TCF7L1 promotes skin tumorigenesis independently of ß-catenin through induction of LCN2. Elife 6:N/A (2017). PubMed: 28467300
Wang X et al. iTRAQ-Based Proteomics Screen identifies LIPOCALIN-2 (LCN-2) as a potential biomarker for colonic lateral-spreading tumors. Sci Rep 6:28600 (2016). WB, IHC . PubMed: 27339395

View all Publications for this product

Customer reviews and Q&As

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1-6 of 6 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (Neutrophils)

Gel Running Conditions

Reduced Denaturing (12% GEL)

Loading amount

25 µg

Specification

Neutrophils

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MME. Louise Chomel

Verified customer

Submitted Jul 30 2021

Application

Western blot

Sample

Human Cell lysate - whole cell (human sebacous gland cells)

Loading amount

60 µg

Specification

human sebacous gland cells

Treatment

13-cis Retinoic acid for 48 hours

Gel Running Conditions

Reduced Denaturing (4- 12% Bis-Tris gel)

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted May 09 2008

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Human skin sections)

Specification

Human skin sections

Fixative

10% Neutral Buffered Formalin

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Trilogy (Cell Marque)

Permeabilization

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 22 2008

Question

thanks will do it.........

Abcam community

Verified customer

Asked on Jan 23 2012

Answer

DISCOUNT CODE: ***********
Expiration date: ***********
I am very pleased to hear you would like to accept our offer and test ab23477 in Rhesus Monkey. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for Rhesus Monkey 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: 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.

Abcam Scientific Support

Answered on Jan 23 2012

Question

I am not clear as to how to go about. I would like to ry the antibody and submit the review . I am not sure how the payments goes through, we order our supplies through the emory express system. Do I have to buy another antibody and then you apply the discount or buy the same antibody and apply the discount.
thanks

Abcam community

Verified customer

Asked on Jan 23 2012

Answer

Thank you for your reply.
As mentioned in the previous email, to take advantage of the testing discount please let me know and I will send you a testing discount code. After you receive the discount code, please follow the steps below:
1 - Place order for ab23477, you will pay full price + shipping and handling.
2 - Test the antibody on rhesus monkey samples.
3 - Write and Abreview on how tha ntibody worked, it can be positive or negative.
4 - Once the Abreview is submitted, then the testing discount code that I originally sent will become active. You can then use that code to receive one free primary antibody of your choice.
I hope that helps explain how the process works, you can find more information in the following link:
https://www.abcam.com/index.htmlpageconfig=resource&rid=11998&viapagetrap=collaborationdiscount
Please let me know if you would like me to send you the testing discount code.
I look forward to your response.

Abcam Scientific Support

Answered on Jan 23 2012

Question

I want to test the antibody Anti-Lipocalin 2 antibody [5G5] (ab23477) for cross reactivity with rhesus monkey and would appreciate if you could provide a sample of the antibody to test which I will be using in a project due. Thanks

Abcam community

Verified customer

Asked on Jan 23 2012

Answer

Thank you very much for your interest in ab23477.
We do not provide sample sizes of our antibodies and since ab23477 has not been tested in rhesus monkey, I can offer a discount off a future purchase if you buy ab23477 now, test it in rhesus monkey and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free PRIMARY ANTIBODY.
If you are interested in this offer, please follow these steps:
1. Reply to this e-mail to let me know that you would like to proceed and test ab(xxx) in rhesus monkey. I will then send a discount code. This code must be issued before purchasing ab23477 so please wait for my reply before ordering.
2. Purchase ab23477 either by phone, fax, or online (www.abcam.com).
3. Test it in rhesus monkey.
4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.
5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any PRIMARY ANTIBODY ordered and the discount code is valid for 4 months after issue.
We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab23477 turns out to be unsuitable for rhesus monkey, you will still receive the discount on your next purchase after your Abreview has been submitted.
Please let me know if you have any questions about this offer and I would be happy to help you further.
The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Abcam Scientific Support

Answered on Jan 23 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Myeloma

Isotype

Light chain type

Research areas

Associated products

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (15)

Customer reviews and Q&As

Western blot abreview for Anti-Lipocalin-2 / NGAL antibody [5G5]

Western blot abreview for Anti-Lipocalin 2 antibody [5G5]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Lipocalin 2 antibody [5G5]