Ki-67/MKI67 Antibody 0.1 ml Proliferation Marker

Click image to view larger
Western Blot: Ki67 Antibody [NB500-170] - Western blot analysis of A431 (A) and Hek293 (B) cell lysate using Ki67 antibody (NB500-170) at 2 ug/ml.
Immunocytochemistry/Immunofluorescence: Ki67 Antibody [NB500-170] - Confocal immunofluorescent analysis of MCF7 cells using Ki67 antibody (NB500-170, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).
Immunohistochemistry-Paraffin: Ki67 Antibody [NB500-170] - Human tonsil.
Immunocytochemistry/Immunofluorescence: Ki67 Antibody [NB500-170] - Ki67 antibody was tested at 1:25 in HeLa cells with FITC (green). Nuclei and actin were counterstained with Dapi (blue) and Phalloidin (red).
Immunocytochemistry/Immunofluorescence: Ki67 Antibody [NB500-170] - Ki67 antibody (1:100) was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red). Image objective 40x.
Immunocytochemistry/Immunofluorescence: Ki67 Antibody [NB500-170] - Ki67 antibody was tested in HeLa cells with Dylight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red). Image objective 40x. A dilution of 1:100 was used.
Download Datasheet
See All Related

Ordering Information: NB500-170

  • Catalog Number
    NB500-170
  • Displayed Format
    Unconjugated
  • Other Formats
  • Size(s)
  • Availability
  • Price
    Shipping
  

Ki-67/MKI67 Antibody Summary

Species Human, Mouse, Rat, Porcine
Tested Applications WB, ICC/IF, IHC, IHC-Fr, IHC-P
Clonality
Polyclonal
Host
Rabbit
Gene
MKI67
Purity
Immunogen affinity purified
Guarantee Plus
All of our products are backed by our 100% guarantee to work for stated and predicted species and applications.
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Ki-67/MKI67 Antibody Details

Isotype
IgG
Marker
Proliferation Marker
Immunogen
A synthetic peptide made to an internal region of human Ki67 (within residues 1550-1700) [Swiss-Prot# P46013]
Localization
Nuclear

Species Reactivity

Human, mouse, rat (pubmed 22521325) and porcine.
Reviewed Species Read 4 rated 4.5
using NB500-170 in the following species:

Publications Read Publications using NB500-170 in the following species:


Applications/Dilutions

Dilutions
  • Western Blot 1-2 ug/ml
  • Immunocytochemistry/Immunofluorescence 1:20-1:100
  • Immunohistochemistry 1:50-1:300
  • Immunohistochemistry-Frozen 1:50-1:300
  • Immunohistochemistry-Paraffin 1:50-1:300
Application Notes
This Ki67 antibody is useful for Immunohistochemistry on frozen and paraffin-embedded sections and Immunocytochemistry/Immunofluorescence. There have been mixed results in Western blot; however, NB500-170 has been used successfully Western Blog reported by a customer review. *Formalin fixed paraffin embedded tissue sections require high temperature antigen unmasking with 10 mM citrate buffer, pH 6.0 prior to immunostaining. This antibody will not work without optimal antigen retrieval. This is probably the most critical step. NOTE: We suggest an incubation period of 30 minutes at room temperature and to use DAB to stain the protein (immunofluorescence may give problems as the protein is nuclear).
Positive Controls
Other Available
Formats
Alexa Fluor (R) 405 Labeled NB500-170AF405
Alexa Fluor (R) 488 Labeled NB500-170AF488
Alexa Fluor (R) 647 Labeled NB500-170AF647
Alexa Fluor (R) 700 Labeled NB500-170AF700
Biotin Labeled NB500-170B
DyLight 405 Labeled NB500-170V
DyLight 488 Labeled NB500-170G
DyLight 550 Labeled NB500-170R
DyLight 650 Labeled NB500-170C
FITC Labeled NB500-170F
Reviewed Applications
Read 3 Review rated 4.5
using
NB500-170 in the following applications:

Publications
Read Publications using
NB500-170 in the following applications:

Contact Information

Product PDFs

Frequently Purchased Together

Reviews

Average Rating: 4.5
Recent Reviews
ICC/IF Tested In Human
Reviewed On: 06/09/2014
Rated: 5

Ki-67/MKI67 NB500-170
Image 1 of 2
Earn rewards points by submitting your review for Ki-67/MKI67 Antibody (NB500-170)

Publications

Ki-67/MKI67 Antibody (NB500-170) has been mentioned in at least 46 publications.
Earn rewards if you have published using Ki-67/MKI67 Antibody (NB500-170).

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Unit Size
0.1 ml
Concentration
1.0 mg/ml
Preservative
0.05% Sodium Azide
Limitations
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.

Bioinformatics

Gene Symbol MKI67
Entrez
Uniprot

Alternate Names for Ki-67/MKI67 Antibody

  • antigen identified by monoclonal Ki-67
  • antigen KI-67
  • KIA
  • proliferation-related Ki-67 antigen
Show More

Related Products by Gene

Background

Originally discovered employing mouse monoclonal antibody against a nuclear antigen from Hodgkin's lymphoma-derived cell line, this non-histone protein was named Ki67 after researcher's location (Gerdes and colleagues), Ki for Kiel University in Germany and 67 referring to the clone number on the 96-well plate. It interacts with KIF15 as well as MKI67IP, and is suggested to be involved in cell cycle regulation. Ki67 is a large protein with expected molecular weight of about 395 kD and has a very complex localization pattern within the nucleus, one which changes during cell cycle progression. Its expression occurs specially during late G1, S, G2 and M phases of the cell cycle, while in cells undergoing G0 phase, Ki67 remains undetectable. Ki67 undergoes phosphorylation/dephosphorylation during mitosis, is susceptible to proteases and its structure implies that its expression is regulated by proteolytic pathways. Ki67 is associated with nucleolar DFC (dense fibrillary component) and its regulation appears to be tightly controlled (estimated half life is 60-90 min, regardless of the cell position in the cell cycle), presumably by precise synthesis and degradation systems involving proteasome, a protease complex. Due to its association with cell divison process, Ki-67 is routinely used as cellular proliferation marker of solid tumors as well as certain hematological malignancies, and a correlation has been demonstrated between Ki-67 index and the histopathological grade of cancers.

Reviews for Ki-67/MKI67 Antibody (NB500-170) (4) 4.54

Read what people are saying who have used Ki-67/MKI67 Antibody (NB500-170).

We have 4 reviews tested in 2 species: Human, Mouse.
We have 3 reviews tested in 3: Immunocytochemistry/Immunofluorescence, Immunocytochemistry, Western Blot.
Average Rating: 4.5
(Based on 4 reviews)

Have you used Ki-67/MKI67 Antibody (NB500-170)? Submit your review and earn rewards points which can be used for merchandise & discounts.

Sort 4 reviews for Ki-67/MKI67 Antibody (NB500-170) by:
Filter by Ratings
Excellent
(2)
Very Good
(2)
All Ratings
Filter by Applications
ICC/IF
(2)
WB
(1)
All Applications
Filter by Species
Human
(3)
Mouse
(1)
All Species

Images Ratings Applications Species Reviewed By Date Details
  Very Good
Human 12/29/2014
Show

Summary

Sample TestedSee PMID 19582877
SpeciesHuman
Immunocytochemistry/Immunofluorescence Ki-67/MKI67 NB500-170
Click to Enlarge
Excellent
ICC/IF Human 06/09/2014
Show

Summary

ApplicationImmunocytochemistry/Immunofluorescence
SpeciesHuman
Special ApplicationsConfocal immunofluorescent analysis of MCF7 cells using Ki67 antibody (NB500-170, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).
FileView PDF
Immunocytochemistry Ki-67/MKI67 NB500-170
Click to Enlarge
Excellent
ICC/IF Human 06/09/2014
Show

Summary

ApplicationImmunocytochemistry
SpeciesHuman
Special ApplicationsConfocal immunofluorescent analysis of MCF7 cells using Ki67 antibody (NB500-170, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).
FileView PDF
  Very Good
WB Mouse 09/30/2010
Show

Summary

ApplicationWestern Blot
Sample TestedWhole cell lysate
SpeciesMouse

Publications for Ki-67/MKI67 Antibody (NB500-170) (46)

Read what people are writing who have used Ki-67/MKI67 Antibody (NB500-170).

We have publications tested in 4 confirmed species: Human, Mouse, Rat, Porcine.

We have publications tested in 4 applications: ICC/IF, IHC, IHC-Fr, IHC-P.

Have you published using Ki-67/MKI67 Antibody (NB500-170)? Submit your review and earn rewards points which can be used for merchandise & discounts.
Filter By Application
ICC/IF
(8)
IHC
(9)
IHC-Fr
(6)
IHC-P
(14)
All Applications
Filter By Species
Human
(12)
Mouse
(16)
Rat
(3)
Porcine
(1)
All Species
Publication Applications Species
Dai G, Freudenberger T, Zipper P et al. Chronic Ultraviolet B Irradiation Causes Loss of Hyaluronic Acid from Mouse Dermis Because of Down-Regulation of Hyaluronic Acid Synthases. Am J Pathol, 2007;171(5):1451-61. 2007 November 1. [PMID: 17982124] (IHC) IHC
Kim SS, Cao L, Baek HJ et al. Impaired skin and mammary gland development and increased gamma-irradiation-induced tumorigenesis in mice carrying a mutation of S1152-ATM phosphorylation site in Brca1. Cancer Res;69(24):9291-300. 2009 Dec 15. [PMID: 19996295]
Zhao Z. Endoplasmic reticulum stress in maternal diabetes-induced cardiac malformations during critical cardiogenesis period. Birth Defects Res B Dev Reprod Toxicol. 2011 Sep 15. [PMID: 21922638] (Mouse) Mouse
Chen F, Loturco J. A method for stable transgenesis of radial glia lineage in rat neocortex by piggyBac mediated transposition. J Neurosci Methods. 2012 Apr 11. [PMID: 22521325] (IHC, Rat) IHC Rat
Ip, M et al. The t10,c12 isomer of conjugated linoleic acid stimulates mammary tumorigenesis in transgenic mice over-expressing erbB2 in the mammary epithelim. Carcinogenesis; 28: 1269 - 1276. 2007 June. [PMID: 17259656]
Gerdes, J et al. Immunobiochemical molecular biologic characterization of the cell proliferation-associated nuclear antigen that is defined by monoclonal antibody Ki-67. Am J Pathol 138: 867-73. 1991 [PMID: 2012175]
Knott, AW et al. Smooth muscle overexpression of IGF-I induces a novel adaptive response to small bowel resection. Am J Physiol Gastrointest Liver Physiol 287: G562-70. 2004 [PMID: 15142831]
Zecchini, V et al. Notch signaling regulates the differentiation of post-mitotic intestinal epithelial cells. Genes Dev 19: 1686-91. 2005 [PMID: 16024658]
Micci, MA et al. Caspase inhibition increases survival of neural stem cells in the gastrointestinal tract. Neurogastroenterol Motil 17: 557-64. 2005 [PMID: 16078945]
Kelly, P et al. THe G12 family of heterotrimeric G proteins promotes breast cancer invasion metastasis. PNAS 103:8173-8178. 2006 [PMID: 16705036]
Ge, R et al. Inhibition of Growth Metastasis of Mouse Mammary Carcinoma by Selective Inhibitor of Transforming Growth Factor-{beta} Type I Receptor Kinase In vivo. Clin, Canc Res 12: 315-4330. 2006 [PMID: 16857807] (IHC-P, Mouse) IHC-P Mouse
Kojima, Kyoko et al. Inactivation of Smad4 Accelerates Kras G12D-Mediated Pancreatic Neoplasia. Cancer Res 67: 8121-8130. 2007 [PMID: 17804724]
Nam, S Y, J S Kim et al. DA-6034, a Derivative of Flavonoid, Prevents Ameliorates Dextran Sulfate Sodium-Induced Colitis Inhibits Colon Carcinogenesis. Experimental Biology Medicine 233: 180-191. 2008 [PMID: 18222973]
Olofsson, P S, L A Soderstrom et al. CD137 Is Expressed in Human Atherosclerosis Promotes Development of Plaque Inflammation in Hypercholesterolemic Mice. Circulation: CIRCULATIONAHA107.699173. 2008 [PMID: 18285570]
ampa VM, Gutierrez-Lanza R, Cerignoli F et al. Notch activates cell cycle reentry progression in quiescent cardiomyocytes. J Cell Biol;183(1):129-141. 2008 [PMID: 18838555]
Gaur S, Chen L, Yen T et al. Preclinical study of the cyclodextrin-polymer conjugate of camptothecin CRLX101 for the treatment of gastric cancer Nanomedicine 2011 Oct 25 [PMID: 22033079] (IHC, Mouse) IHC Mouse
McCaskey SJ, Rondini EA, Langohr IM, Fenton JI. Differential effects of energy balance on experimentally-induced colitis World J Gastroenterol 2012 Feb 21 [PMID: 22363133] (IHC, Mouse) IHC Mouse
Rock K, Meusch M, Fuchs N et al. Estradiol Protects Dermal Hyaluronan/Versican Matrix during Photoaging by Release of Epidermal Growth Factor from Keratinocytes J Biol Chem 2012 Jun 8 [PMID: 22493503] (IHC-Fr, Mouse) IHC-Fr Mouse
Wu X, Zhang Q, Wang X et al. Survivin Is Required for Beta-Cell Mass Expansion in the Pancreatic Duct-Ligated Mouse Model plos One 2012 [PMID: 22870272] (IHC, Mouse) IHC Mouse
Rauhavirta T, Oittinen M, Kivisto R et al. Are Transglutaminase 2 Inhibitors Able to Reduce Gliadin-Induced Toxicity Related to Celiac Disease? A Proof-of-Concept Study J Clin Immunol 2012 Aug 10 [PMID: 22878839] (IHC-Fr, Human) IHC-Fr Human
Zhang Y, Srinivasan B, Xing C, Lu J. A New Chalcone Derivative (E)-3-(4-Methoxyphenyl)-2-methyl-1-(3,4,5-trimethoxyphenyl)prop-2-en-1-one Suppresses Prostate Cancer Involving p53-mediated Cell Cycle Arrests and Apoptosis Anticancer Res 2012 Sep [PMID: 22993307]
Lee NJ, Choi DY, Song JK et al. Deficiency of C-C chemokine receptor 5 suppresses tumor development via inactivation of NF-{kgreen}B and inhibition of monocyte chemoattractant protein-1 in urethane-induced lung tumor model Carcinogenesis 2012 Oct 5 [PMID: 22907530] (IHC, Mouse) IHC Mouse
Tao YM, Huang JL, Zeng S et al. BTB/POZ domain-containing protein 7: Epithelial-mesenchymal transition promoter and prognostic biomarker of hepatocellular carcinoma. Hepatology 2013 Jan 16 [PMID: 23325674] (IHC-P, Human, Mouse) IHC-P Human, Mouse
Imamura F, Greer CA. Pax6 regulates Tbr1 and Tbr2 expressions in olfactory bulb mitral cells. Mol Cell Neurosci 2013 Jan 22 [PMID: 23353076] (IHC-Fr, ICC/IF, Mouse) ICC/IF, IHC-Fr Mouse
Mildner M, Hacker S, Haider T et al. Secretome of Peripheral Blood Mononuclear Cells Enhances Wound Healing PLoS One 2013 [PMID: 23533667] (IHC-P, ICC/IF, Human) ICC/IF, IHC-P Human
Saggar JK, Fung AS, Patel KJ, Tannock IFet al. Use of molecular biomarkers to quantify the spatial distribution of effects of anticancer drugs in solid tumors. Mol Cancer Ther 2013 Apr [PMID: 23348047] (IHC, ICC/IF, Human) ICC/IF, IHC Human
Kirby ED, Muroy SE, Sun WG et al. Acute stress enhances adult rat hippocampal neurogenesis and activation of newborn neurons via secreted astrocytic FGF2. Elife 2013 Apr 16 [PMID: 23599891] (IHC-Fr, ICC/IF, Rat) ICC/IF, IHC-Fr Rat
Biddlestone-Thorpe L, Sajjad M, Rosenberg E et al. ATM Kinase Inhibition Preferentially Sensitizes p53-Mutant Glioma to Ionizing Radiation. Clin Cancer Res 2013 Jun 15 [PMID: 23620409] (IHC-Fr, ICC/IF, Human) ICC/IF, IHC-Fr Human
Maeda N, Kokai Y, Ohtani S et al. Anti-tumor effect of orally administered spinach glycolipid fraction on implanted cancer cells, colon-26, in mice. Lipids 2008 Aug [PMID: 18594894] (IHC-P, Mouse) IHC-P Mouse
Spiegelberg L, Braks J, Djasim U et al. Effects of hyperbaric oxygen therapy on the viability of irradiated soft head and neck tissues in mice. Oral Dis 2013 Jul 8 [PMID: 23865954] (IHC-P, Mouse) IHC-P Mouse
Xiang L, Gilkes DM, Chaturvedi P, Semenza GL et al. Ganetespib blocks HIF-1 activity and inhibits tumor growth, vascularization, stem cell maintenance, invasion, and metastasis in orthotopic mouse models of triple-negative breast cancer. J Mol Med (Berl). 2013 Nov 20 [PMID: 24248265]
Liu Y, Sanchez-Tillo E, Lu X et al. Rb1 family mutation is sufficient for sarcoma initiation. Nat Commun. 2013 Oct 23 [PMID: 24150016] (ICC/IF, Mouse) ICC/IF Mouse
Niedworok C, Rock K, Kretschmer I et al. Inhibitory Role of the Small Leucine-Rich Proteoglycan Biglycan in Bladder Cancer. PLoS One. 2013 Nov 6 [PMID: 24223213] (IHC, Human) IHC Human
Saggar JK, Tannock IF. Activity of the hypoxia-activated pro-drug TH-302 in hypoxic and perivascular regions of solid tumors and its potential to enhance therapeutic effects of chemotherapy. Int J Cancer. 2013 Nov 8 [PMID: 24338277] (IHC-Fr, ICC/IF, Human) ICC/IF, IHC-Fr Human
Kwon S, Kim MS, Lee ES, Park JK. A quantum dot-based microfluidic multi-window platform for quantifying the biomarkers of breast cancer cells. Integr Biol (Camb) 2014 Mar 5 [PMID: 24599518] (IHC-P, ICC/IF, Human) ICC/IF, IHC-P Human
Gou HF, Huang J, Shi HS, Wang YS. Chemo-immunotherapy with oxaliplatin and interleukin-7 inhibits colon cancer metastasis in mice. PLoS ONE 2014 Jan 27 [PMID: 24465710] (IHC-P, Mouse) IHC-P Mouse
Sano M, Sasaki T, Hirakawa S et al. Lymphangiogenesis and angiogenesis in abdominal aortic aneurysm. PLoS ONE 3/21/2014 [PMID: 24651519] (IHC-P, Human) IHC-P Human
Sherman-Baust CA, Kuhn E, Valle BL et al. A genetically engineered ovarian cancer mouse model based on fallopian tube transformation mimics human high-grade serous carcinoma development. J. Pathol. 3/20/2014 [PMID: 24652535] (IHC-P, Mouse) IHC-P Mouse
Takano N, Sarfraz Y, Gilkes DM et al. Decreased Expression of Cystathionine b-Synthase Promotes Glioma Tumorigenesis. Mol. Cancer Res. 2014 Jul 03 [PMID: 24994751] (IHC-P, Human)

Details:
Ki67 antibody used for IHC-P on tumor xenografts derived from injection of parental U87-MG cells and shCBS#07 subclone (Fig 1E, Fig 3D).
IHC-P Human
Hu H, Takano N, Xiang L et al. Hypoxia-inducible factors enhance glutamate signaling in cancer cells. Oncotarget 2014 Oct 15 [PMID: 25326682] (IHC-P, Human) IHC-P Human
Xie H, Cui Z, Wang L et al. PDGF-BB secreted by preosteoclasts induces angiogenesis during coupling with osteogenesis. Nat. Med. 2014 Oct 05 [PMID: 25282358] (IHC, Mouse) IHC Mouse
Uegaki T, Taniguchi F, Nakamura K et al. Inhibitor of apoptosis proteins (IAPs) may be effective therapeutic targets for treating endometriosis Hum. Reprod. 2014 Nov 05 [PMID: 25376458] (IHC-P, Mouse) IHC-P Mouse
Kawanishi Y, Nakasa T, Shoji T et al. Intra-articular injection of synthetic microRNA-210 accelerates avascular meniscal healing in rat medial meniscal injured model. Arthritis Res. Ther. 2014 Nov 28 [PMID: 25430980] (IHC-P, Rat)

Details:
Ki67 antibody used for IHC-P in Spraque Dawley rats subjected to intra-articular injection of double-stranded miR-210 (for control group using control dsRNA) with atelocollagen administered immediately after injury (injured meniscal healing in a rat model) - 4%PFA fixation, 4um sections, blocking with horse serum, Ki67 antibody diluted at 1:100 (Figure 4).
IHC-P Rat
Nakagawa Y, Muneta T, Kondo S et al. Synovial mesenchymal stem cells promote healing after meniscal repair in microminipigs Osteoarthr. Cartil. 2015 Feb 13 [PMID: 25683149] (IHC-P, Porcine) IHC-P Porcine
Smith Corey L, Matheson Timothy D, Trombly Daniel J et al. A separable domain of the p150 subunit of human chromatin assembly factor-1 promotes protein and chromosome associations with nucleoli. Mol Biol Cell. 2014 Sep 15 [PMID: 25057015] (Human) Human
Mertens Marianne E, Hermann Alina, Buhren Anne et al. Iron Oxide-labeled Collagen Scaffolds for Non-invasive MR Imaging in Tissue Engineering. Adv Funct Mater. 2014 Feb 12 [PMID: 24569840]

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

Ask a Scientist

Submit your question on below.
During business hours, we will respond to your email within 24 hours. For any questions submitted on the weekend, a response will be received on Monday.
For immediate assistance during business hours M- F (excluding major holidays), please contact us.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.

FAQs for Ki-67/MKI67 (33)

  1. Could you please let us know if you have the antibody which may have cross-reactivity to hamster tissue in IHC(paraffin or frozen)? Also, please tell us reference information using your products to hamster tissue.
    • Thank you for your patience. The only information I was able to gain is that the protein has 61% homology with the chinese hamster. Our lab only recommends use if the % homology is 85% or above so I would not recommend our Ki67 antibodies for your customer. However if they are willing to still test this they would become eligable for our Innovators Reward Program in which they would gain a 50% refund and a 50% discount on a future purchase. All you would need to do is email us at innovators@novusbio.com. Here is the link to all of our Ki67 antibodies.
  2. What is the IF protocol which was used to obtain the IF image on the datasheet?
    • Our lab used our general ICC protocol to obtain the image seen on our website.
  3. In the immunohistochemistry paraffin section protocol....the PBS buffer....should it be at certain pH?
    • For IHC application, different labs cites the pH of PBS buffer in the range of 7.1 - 7.6 but most commonly used pH is pH 7.4 (this is what we use in our lab). You may use this PBS (pH 7.4) for making permeablization buffer, antibody diluent buffers and for making wash buffer. For more on the protocol that we use in our lab and for IHC-P troubleshooting suggestions, you may visit: IHC-P Protocol and IHC-P Troubleshooting.
  4. I was wondering what does Format 7C mean as far as anitbodies go?
    • 7C is a fluorophore - it is useful for FLOW. The details of the fluor are (A=425, E=500)
  5. What epitope does this monoclonal antibody recognize?
    • R&D Systems does not epitope map its antibodies. The antibody is generated using the entire immunogen listed on the datasheet. If a peptide immunogen is used, this will be indicated on the datasheet.
  6. What epitope does this polyclonal antibody recognize?
    • Polyclonal antibodies generally recognize multiple epitopes because they are generated using the entire immunogen stated on the product datasheet.
  7. What are R&D Systems matched antibody pairs?
    • R&D Systems matched antibody pairs assist in the development of an ELISA. R&D Systems has demonstrated that the paired antibodies can be used in a sandwich immunoassay to recognize the recombinant protein. We recommend that the investigator have expertise in immunoassay development before attempting to use these products. Each investigator must empirically determine the optimal concentrations for the capture and detection antibodies. The amount of antibodies supplied are not normalized to develop the same number of plates. R&D Systems recombinant cytokines are not calibrated by ELISA and therefore must be mass calibrated before use. The DuoSet ELISA Development Kit product line includes the antibodies, standard, enzyme, and protocol necessary for running an immunoassay, and can often be a better value than purchasing antibody pairs and protein standard.
  8. Can I use your antibodies for a matched pair ELISA with a standard from another company?
    • Yes, but there are potential issues to consider. There can be differences in immunological recognition or the binding between antibody and recombinant protein. This can happen if there is a difference in the folding or sequence of the recombinant protein used as the standard versus the recombinant protein used as the immunogen for the antibody. Protein folding or sequence differences in the antibody-binding region can lead to poor (or no) recognition of the standard.
  9. Can I use your antibodies for a matched pair ELISA with a standard from another company?
    • Yes, but there are potential issues to consider. There can be differences in immunological recognition or the binding between antibody and recombinant protein. This can happen if there is a difference in the folding or sequence of the recombinant protein used as the standard versus the recombinant protein used as the immunogen for the antibody. Protein folding or sequence differences in the antibody-binding region can lead to poor (or no) recognition of the standard.
  10. What grade BSA is recommend for use in ELISA and ELISpot Development Systems?
    • The grade of the BSA used has been found to be a critical component for running a successful ELISA. BSA with a minimum purity of 98% is recommended. R&D Systems recommends using the same BSAs used in development of the DuoSet; Catalog # DY995 Reagent Diluent Concentrate 2, Catalog # DY997 Reagent Diluent Concentrate 1. Millipore Probumin® Diagnostic Grade BSA (Millipore, Catalog # 82-045) is another example.
  11. What grade BSA is recommend for use in ELISA and ELISpot Development Systems?
    • The grade of the BSA used has been found to be a critical component for running a successful ELISA. BSA with a minimum purity of 98% is recommended. R&D Systems recommends using the same BSAs used in development of the DuoSet; Catalog # DY995 Reagent Diluent Concentrate 2, Catalog # DY997 Reagent Diluent Concentrate 1. Millipore Probumin® Diagnostic Grade BSA (Millipore, Catalog # 82-045) is another example.
  12. What is a direct ELISA?
    • In a direct ELISA, a plate is coated with the analyte of interest and a labeled detection antibody is used to verify the presence of the analyte.The direct ELISA may use a colorimetric, chemiluminescent, or fluorescent reporter.
  13. What is a direct ELISA?
    • In a direct ELISA, a plate is coated with the analyte of interest and a labeled detection antibody is used to verify the presence of the analyte.The direct ELISA may use a colorimetric, chemiluminescent, or fluorescent reporter.
  14. What is a sandwich ELISA?
    • A sandwich ELISA uses an immobilized capture antibody specific for the analyte of interest in a sample. After the analyte is bound to the immobilized antibody, a labeled secondary antibody specific for the analyte is used for detection. The analyte is "sandwiched" between the two antibodies. The sandwich ELISA is extremely sensitive, and the values obtained are quantitative when compared with a standard curve.
  15. What is a sandwich ELISA?
    • A sandwich ELISA uses an immobilized capture antibody specific for the analyte of interest in a sample. After the analyte is bound to the immobilized antibody, a labeled secondary antibody specific for the analyte is used for detection. The analyte is "sandwiched" between the two antibodies. The sandwich ELISA is extremely sensitive, and the values obtained are quantitative when compared with a standard curve.
  16. What is the difference between AB###, AF###, BAF###, MAB###, and BAM### antibody catalog number designations?
    • AB### designated antibodies are polyclonal antibodies that are purified using Protein A, Protein G or Ion Exchange chromatography. Consequently, the resulting purified antibody is a total IgG fraction and may contain IgG not specific for the analyte of interest. AF### designated antibodies, are antigen affinity-purified and then further purified by ion exchange or size exclusion. Therefore, AF### antibodies are IgG specific only to the antigen. Antibodies that have the designation MAB### are monoclonal antibodies. BAF### and BAM### designated antibodies are the biotinylated versions of the AF### and MAB### designated antibodies, respectively.
  17. What is the difference between AB###, AF###, BAF###, MAB###, and BAM### antibody catalog number designations?
    • AB### designated antibodies are polyclonal antibodies that are purified using Protein A, Protein G or Ion Exchange chromatography. Consequently, the resulting purified antibody is a total IgG fraction and may contain IgG not specific for the analyte of interest. AF### designated antibodies, are antigen affinity-purified and then further purified by ion exchange or size exclusion. Therefore, AF### antibodies are IgG specific only to the antigen. Antibodies that have the designation MAB### are monoclonal antibodies. BAF### and BAM### designated antibodies are the biotinylated versions of the AF### and MAB### designated antibodies, respectively.
  18. What is the rationale for using trehalose to stabilize proteins?
    • Trehalose is an effective sugar for stabilizing proteins against damage caused by freezing. It can also make the protein more resistant to moisture gain when lyophilized, resulting in a product that is less likely to precipitate when reconstituted. In addition, it has been used in approved parenteral therapeutics
  19. What is the rationale for using trehalose to stabilize proteins?
    • Trehalose is an effective sugar for stabilizing proteins against damage caused by freezing. It can also make the protein more resistant to moisture gain when lyophilized, resulting in a product that is less likely to precipitate when reconstituted. In addition, it has been used in approved parenteral therapeutics
  20. Will trehalose affect my conjugation reaction?
    • It is possible that the presence of trehalose will interfere in the successful conjugation of a protein. This will depend on the method used, and the customer should investigate this potential prior to purchasing the product.
  21. Will trehalose affect my conjugation reaction?
    • It is possible that the presence of trehalose will interfere in the successful conjugation of a protein. This will depend on the method used, and the customer should investigate this potential prior to purchasing the product.
  22. Will trehalose affect the performance of the protein or antibody in my specific application?
    • We have seen no adverse effect in our bioassays or other approved applications. However, customers are advised to run a control in their assay to determine if the concentration of trelahose in the protein or antibody formulation has any adverse effects.
  23. Will trehalose affect the performance of the protein or antibody in my specific application?
    • We have seen no adverse effect in our bioassays or other approved applications. However, customers are advised to run a control in their assay to determine if the concentration of trelahose in the protein or antibody formulation has any adverse effects.
  24. Will trehalose included in the formulation affect the animal if it is injected?
    • Trehalose is unlikely to have an effect in vivo. It has been approved as an excipient for use in human injectable drugs.The trehalose used by R&D Systems is derived from Saccharomyces cerevisiae and is determined to be at minimum 98.5% pure by HPAE.
  25. Will trehalose included in the formulation affect the animal if it is injected?
    • Trehalose is unlikely to have an effect in vivo. It has been approved as an excipient for use in human injectable drugs.The trehalose used by R&D Systems is derived from Saccharomyces cerevisiae and is determined to be at minimum 98.5% pure by HPAE.
  26. What are R&D Systems matched antibody pairs?
    • R&D Systems matched antibody pairs assist in the development of an ELISA. R&D Systems has demonstrated that the paired antibodies can be used in a sandwich immunoassay to recognize the recombinant protein. We recommend that the investigator have expertise in immunoassay development before attempting to use these products. Each investigator must empirically determine the optimal concentrations for the capture and detection antibodies. The amount of antibodies supplied are not normalized to develop the same number of plates. R&D Systems recombinant cytokines are not calibrated by ELISA and therefore must be mass calibrated before use. The DuoSet ELISA Development Kit product line includes the antibodies, standard, enzyme, and protocol necessary for running an immunoassay, and can often be a better value than purchasing antibody pairs and protein standard.
  27. What is the recommended protocol for reconstituting and aliquotting lyophilized proteins and antibodies?<br>
    • Following these guidelines for reconstituting lyophilized material will help ensure complete recovery of the protein or antibody.

      1) Tap or briefly centrifuge the vial before opening to dislodge any lyophilized material that may be dispersed on the wall or cap of the vial.

      2) Use the buffer and stock concentration recommended in the product datasheet. If you want a stock concentration that is higher than the one recommended, contact Technical Service for specific recommendations.

      3) For optimal recovery, reconstitute using room temperature buffer.

      4) After adding the buffer, re-cap the vial and invert gently by hand or place on a slow rocking platform. This will allow the reconstitution buffer to coat all the surfaces inside the vial. Do not mix by vortexing or by pipetting the material up and down.

      5) Allow the vial to sit at room temperature with gentle agitation for at least 15 minutes before aliquotting or using.

      6) Store the reconstituted protein in polypropylene or siliconized tubes. If aliquotting, it is recommended that aliquots be no smaller than 10 μL. In addition, avoid repeated freeze/thaw cycles.

  28. What is the recommended protocol for reconstituting and aliquotting lyophilized proteins and antibodies?<br>
    • Following these guidelines for reconstituting lyophilized material will help ensure complete recovery of the protein or antibody.

      1) Tap or briefly centrifuge the vial before opening to dislodge any lyophilized material that may be dispersed on the wall or cap of the vial.

      2) Use the buffer and stock concentration recommended in the product datasheet. If you want a stock concentration that is higher than the one recommended, contact Technical Service for specific recommendations.

      3) For optimal recovery, reconstitute using room temperature buffer.

      4) After adding the buffer, re-cap the vial and invert gently by hand or place on a slow rocking platform. This will allow the reconstitution buffer to coat all the surfaces inside the vial. Do not mix by vortexing or by pipetting the material up and down.

      5) Allow the vial to sit at room temperature with gentle agitation for at least 15 minutes before aliquotting or using.

      6) Store the reconstituted protein in polypropylene or siliconized tubes. If aliquotting, it is recommended that aliquots be no smaller than 10 μL. In addition, avoid repeated freeze/thaw cycles.

  29. Can this antibody be used on frozen or paraffin-embedded&nbsp; sections?
    • R&D Systems will provide support for an antibody that is validated for the IHC/ICC application in both frozen and paraffin-embedded sections unless otherwise specified on the datasheet. Should the product fail to work after contacting us for assistance, R&D Systems will offer a product credit toward a future purchase.

      If the datasheet does not have an IHC/ICC claim, we do not have any in-house or collaborative data available to support this application and we cannot guarantee results. It is up to the user to decide if they wish to use the antibody in their application.

      Customers may use the Citations tab on the product-specific web page or contact Technical Service to check for references using a product in an application or with a cell type R&D Systems has not tested in-house.
  30. Can this antibody be used on frozen or paraffin-embedded&nbsp; sections?
    • R&D Systems will provide support for an antibody that is validated for the IHC/ICC application in both frozen and paraffin-embedded sections unless otherwise specified on the datasheet. Should the product fail to work after contacting us for assistance, R&D Systems will offer a product credit toward a future purchase.

      If the datasheet does not have an IHC/ICC claim, we do not have any in-house or collaborative data available to support this application and we cannot guarantee results. It is up to the user to decide if they wish to use the antibody in their application.

      Customers may use the Citations tab on the product-specific web page or contact Technical Service to check for references using a product in an application or with a cell type R&D Systems has not tested in-house.
  31. Can this antibody be used on frozen or paraffin-embedded&nbsp; sections?
    • R&D Systems will provide support for an antibody that is validated for the IHC/ICC application in both frozen and paraffin-embedded sections unless otherwise specified on the datasheet. Should the product fail to work after contacting us for assistance, R&D Systems will offer a product credit toward a future purchase.

      If the datasheet does not have an IHC/ICC claim, we do not have any in-house or collaborative data available to support this application and we cannot guarantee results. It is up to the user to decide if they wish to use the antibody in their application.

      Customers may use the Citations tab on the product-specific web page or contact Technical Service to check for references using a product in an application or with a cell type R&D Systems has not tested in-house.
  32. Has a product ever been used for an application that is not listed on the datasheet?
    • If a specific application is not listed on the datasheet, it may mean that this product has not been tested in this application, or it may mean that in-house testing in this application did not meet R&D Systems’specifications. Please check our Citations tab to see if other researchers have published using your application, sample type and/or species. If you would like more information on whether or not an application has been tested, please contact Technical Service at (800) 343-7475.

  33. Has a product ever been used for an application that is not listed on the datasheet?
    • If a specific application is not listed on the datasheet, it may mean that this product has not been tested in this application, or it may mean that in-house testing in this application did not meet R&D Systems’specifications. Please check our Citations tab to see if other researchers have published using your application, sample type and/or species. If you would like more information on whether or not an application has been tested, please contact Technical Service at (800) 343-7475.