Carbonic Anhydrase IX/CA9 Antibody (NB100-417)

Images

 
Immunohistochemistry: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis using the Biotin conjugate. Staining of Renal carcinoma tissue.
Immunohistochemistry-Paraffin: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - IHC analysis of a formain fixed paraffin embedded (FFPE) tissue section of human breast cancer using CAIX antibody at 1:1000 dilution. The ...read more
Immunohistochemistry-Frozen: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis of human colon carcinoma, xenografted in mice. Image courtesy of an anonymous product review.
Western Blot: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis in 1) HeLa, 2) MDA-MB-231 and 3) A549 whole cell lysates.
Immunocytochemistry/ Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis of HRP conjugate of NB100-417. HEK 293 cells using NB100-417. Panel A shows CAIX-transfected cells (epitope-unmasked ...read more
Western Blot: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis using the HRP conjugate. Analysis in transfected HEK cell lysate. Rabbit IgG was used as a negative control.
Immunocytochemistry/ Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis using the DyLight 488 conjugate of NB100-417. Staining of Carbonic Anhydrase IX (red) in human glioma U87 cells. DAPI ...read more
Western Blot: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Anaysis in transfected HEK cell lysate using NB 100-417. Rabbit IgG was used as a negative control.
Western Blot: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis on rat renal cortex.
Western Blot: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis on various human cell lysates. Image from verified customer review.
Western Blot: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Detection of HRP conjugated carbonic anhydrase IX in transfected HEK cell lysate. Rabbit IgG was used as a negative control.
Immunocytochemistry/ Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis of Biotin conjugated antibody on HEK 293 cells. Panel A shows CAIX-transfected cells (epitope-unmasked with SDS) and ...read more
Immunocytochemistry/ Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Immunofluorescence of human RCC tumor cryosections using NB100-417 (Panel A). Panel B shows staining with normal rabbit serum.
Immunocytochemistry/ Immunofluorescence: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Analysis of Biotin conjugate of NB100-417. Human RCC tumor cryosections using NB100-417 (Panel A). Panel B shows staining with ...read more
Simple Western: Carbonic Anhydrase IX/CA9 Antibody [NB100-417] - Simple Western lane view shows a specific band for CAIX in 0.1 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Ca, PlSpecies Glossary
Applications WB, Simple Western, ELISA, Flow, GS, ICC/IF, IHC, IHC-Fr, IHC-P, ChIP
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.0 mg/ml

Carbonic Anhydrase IX/CA9 Antibody Summary

Immunogen
A synthetic peptide made to a C-terminal sequence of human Carbonic Anhydrase IX (within residues 400-459) [UniProt Q16790]
Localization
Membrane.
Marker
Hypoxia Marker
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
CA9
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:2000
  • Simple Western 1:50
  • ELISA
  • Flow Cytometry
  • Gel Super Shift Assays
  • Immunocytochemistry/Immunofluorescence 1:1000
  • Immunohistochemistry 1:1000
  • Immunohistochemistry-Frozen 1:1000
  • Immunohistochemistry-Paraffin 1:1000
  • Chromatin Immunoprecipitation (ChIP)
Application Notes
This Carbonic Anhydrase IX antibody is useful for Western blot, Immunofluorescence/Immunocytochemistry, and Immunohistochemistry. Gel Super Shift Assays was reported in scientific literature. In Western blot a band is observed approx. 53 kDa. Use in ELISA reported in scientific literature (PMID 19963243). Use in chromatin immunoprecipitation reported in multiple pieces of scientific literature.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
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.
Theoretical MW
55 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.
Positive Control
HeLa Lysate (NB800-PC1)
Carbonic Anhydrase IX/CA9 Lysate (NBL1-08619)
Control Peptide
Carbonic Anhydrase IX/CA9 Peptide (NB100-417PEP)
Reviewed Applications
Read 3 Reviews rated 4.7
using
NB100-417 in the following applications:

Publications
Read Publications using
NB100-417 in the following applications:

Reactivity Notes

Human, dog, rat and mouse (PMID 22842475). Plant reactivity reported in scientific literature (PMID: 19278636)

Packaging, Storage & Formulations

Storage
Store at -20C. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.025% Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

Alternate Names for Carbonic Anhydrase IX/CA9 Antibody

  • CA9
  • CA-IX
  • CAIXcarbonic anhydrase 9
  • Carbonate dehydratase IX
  • Carbonic Anhydrase IX
  • carbonic anhydrase IXpMW1
  • carbonic dehydratase
  • EC 4.2.1.1
  • G250
  • Membrane antigen MN
  • MN
  • MNRCC-associated antigen G250
  • P54/58N
  • RCC
  • RCC-associated protein G250
  • Renal cell carcinoma-associated antigen G250

Background

Carbonic anhydrase IX (CAIX) is a member of the Carbonic anhydrase family which are enzymes that assists rapid interconversion of carbon dioxide and water into carbonic acid, protons, and bicarbonate ions. They are abundant in all mammalian tissues and because of their functionality, they have become an important diagnostic marker for various cancers, most notably renal cell carcinoma (RCC). Carbonic Anhydrase IX (CA IX) is one of the most hypoxically inducible genes because of its stability and the location of the expressed protein within the membrane. Carbonic anhydrases have a widespread role in regulating pH in normal tissues, by regulating hydrogen ion (H+) flux. The pH is important in cell death under hypoxia, thus a blockade of CA IX (CA9) results in increased cell death under hypoxia. Therefore, CA IX has become a reliable histochemical marker of hypoxia.

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.

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Publications for Carbonic Anhydrase IX/CA9 Antibody (NB100-417)(139)

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

We have publications tested in 9 applications: ChIP, ELISA, EMSA, ICC/IF, IHC, IHC-FR, IHC-Fr, IHC-P, WB.


Filter By Application
ChIP
(2)
ELISA
(1)
EMSA
(1)
ICC/IF
(10)
IHC
(44)
IHC-FR
(1)
IHC-Fr
(4)
IHC-P
(41)
WB
(18)
All Applications
Filter By Species
Human
(88)
Mouse
(19)
Rat
(4)
Plant
(1)
All Species
Showing Publications 1 - 10 of 139. Show All 139 Publications.
Publications using NB100-417 Applications Species
Klameth L, Rath B, Hochmaier M et al. Small cell lung cancer: model of circulating tumor cell tumorospheres in chemoresistance Sci Rep 2017 Jul 13 [PMID: 28706293] (IHC-P, Human) IHC-P Human
Zhang Y, Sun M, Huang G et al. Maintenance of antiangiogenic and antitumor effects by orally active low-dose capecitabine for long-term cancer therapy. Proc. Natl. Acad. Sci. U.S.A. 2017 Jun 27 [PMID: 28607065]
Takenaka M, Suzuki N, Mori M et al. Iron regulatory protein 2 in ovarian endometrial cysts. Biochem. Biophys. Res. Commun. Apr 24 2017 [PMID: 28450115]
Bousquet G, El Bouchtaoui M, Sophie T et al. Targeting autophagic cancer stem-cells to reverse chemoresistance in human triple negative breast cancer. Oncotarget. 2017 May 23 [PMID: 28445132] (ICC/IF, Human) ICC/IF Human
Himmels P, Paredes I, Adler H et al. Motor neurons control blood vessel patterning in the developing spinal cord. Nat Commun. Mar 6 2017 [PMID: 28262664] (IHC-FR, Mouse) IHC-FR Mouse
Fraga A, Ribeiro R, Coelho A et al. Genetic polymorphisms in key hypoxia-regulated downstream molecules and phenotypic correlation in prostate cancer. BMC Urol 2017 Jan 31 [PMID: 28143503] (Human) Human
Bulusu V, Tumanov S, Michalopoulou E et al. Acetate Recapturing by Nuclear Acetyl-CoA Synthetase 2 Prevents Loss of Histone Acetylation during Oxygen and Serum Limitation Cell Rep Jan 17 2017 [PMID: 28099844] (IHC, Human) IHC Human
Tsuji T, Kurokawa Y, Chiche J et al. Dissecting the process of activation of cancer-promoting zinc-requiring ectoenzymes by zinc metalation mediated by ZnT transporters J. Biol. Chem Dec 27 2016 [PMID: 28028180] (WB) WB
Oh ET, Kim JW, Kim JM et al. NQO1 inhibits proteasome-mediated degradation of HIF-1a. Nat Commun. Dec 14 2016 [PMID: 27966538] (IHC, Mouse) IHC Mouse
Frost J, Galdeano C, Soares P et al. Potent and selective chemical probe of hypoxic signalling downstream of HIF-alpha hydroxylation via VHL inhibition. Nat Commun. Nov 4 2016 [PMID: 27811928] (WB) WB
Show All 139 Publications.

Reviews for Carbonic Anhydrase IX/CA9 Antibody (NB100-417) (3) 4.73

Average Rating: 4.7
(Based on 3 reviews)
We have 3 reviews tested in 1 species: Human.
We have 3 reviews tested in 2 application: WB, ICC.

Reviews using NB100-417:
Filter by Applications
WB
(2)
ICC
(1)
All Applications
Filter by Species
Human
(3)
All Species
Images Ratings Applications Species Date Details
 
reviewed by:
rajesha rupaimoole
WB Human 12/10/2014
View

Summary

ApplicationWestern Blot
Sample TestedA2780 ovarian cancer cells
SpeciesHuman

Blocking

Blocking Details5% BSA

Details

Detection NotesECL
Western Blot Carbonic Anhydrase IX/CA9 NB100-417
Enlarge
reviewed by:
Xiuquan Luo
WB Human 07/21/2014
View

Summary

ApplicationWestern Blot
Sample Tested20 ug whole cell lysate
SpeciesHuman

Blocking

Blocking Details1xPBST with 5% non-fat milk, RT 1 hour

Primary Anitbody

Dilution Ratio1xpBST with 5% non-fat milk, overnight 4C, 1:1000 dilution

Secondary Antibody

Secondary DescriptionGoat anti-rabbit IgG-HRP
Secondary Manufacturer Cat#sc-2004
Secondary Concentration1:2000 dilution

Details

Detection NotesECL, exposured for 3 min
Immunocytochemistry Carbonic Anhydrase IX/CA9 NB100-417
Enlarge
reviewed by:
Sonia Rocha
ICC Human 12/21/2011
View

Summary

ApplicationImmunocytochemistry
Sample TestedHuman cells
SpeciesHuman
CommentsReally nice results using microscopy.

Blocking

Blocking DetailsBlocking Buffer: 1% Donkey serum PBS, Blocking Time: 30 minutes, Blocking Temp: 20 degrees Celsius

Primary Anitbody

Dilution RatioIncubation Dilution Buffer: PBS/1% Donkey serum, Dilution Ratio: 1:250, Incubation Time: 1 hour, Incubation Temp: 20 C

Secondary Antibody

Secondary DescriptionSecondary Ab: anti-rabbit alexa 488, Secondary Ab Dilution: 1:500

Details

Detection NotesDetection Visualization Method: fluorescence at 488, Negative Control: secondary and primary only
Fixation DetailsSample Preparation: PFA fixed

Comments

CommentsReally nice results using microscopy.

Product General Protocols

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for Carbonic Anhydrase IX/CA9 Antibody (NB100-417). (Showing 1 - 8 of 8 FAQs).

  1. How many units do we need to purchase in order to stain 150 formalin fixed, paraffin embedded kidney biopsies? It depends on the amount used per slide, so could you please confirm how much antibody do you use per slide in IHC-P? Also, is the ab compatible with ENVISION FLEX+, HIGH pH, FOR AS?
    • Unfortunately there is no number that I can provide to you as it is impossible to know for sure how many slides you will be able to stain with a vial. This will depend on your experimental conditions, your samples and how highly expressed the protein is, and what dilution you end up choosing. This antibody is recommended to be used at a 1:1000 dilution for IHC staining as this has worked well for our lab. You should order a single vial and test to determine what amount will be best for your assay rather than order too much or too little without first working with the antibody. As for your second question, that is not something that our lab has tested with so I am unsure if it is compatible or not. It has a fairly normal range pH in the buffer (7-8) so most likely the functions best around that range and not at a high pH.
  2. Could you please let me know the immunogen peptide location for this Ab?
    • The immunogen used to create this antibody was made to a cytoplasmic area of the protein.
  3. I'm going to use your CA-9 Antibody for paraffin-Immunohistochemistry. Do I have to do any special pretreatments?
    • Here is our protocol specific for this product. However, we recommend antigen retrieval to ensure the epitope is not masked, and we also recommend that you add a permeablizing agent to access the epitope in the nucleus.
  4. We dilute our antibodies in 5% milk TBS-Tween and usually freeze-thaw them multiple times. Is this possible with this antibody when diluted?
    • Freezing is not recommended for NB100-417 under any circumstances. Unfortunately, the antibody would no longer be covered by our guarantee if it was stored at a temperature that did not match our storage instructions. Of course, you are free to test your proposed conditions if you would like, but we would not be able to accept responsibility if the antibody was damaged by this.
  5. Is this antibody is carrier-free? If not, do you have a carrier-free version?
    • NB100-417 is carrier protein free, but does contain 0.1% sodium azide. The buffer formulation is: Tris-citrate/phosphate, pH 7-8 with 0.1% sodium azide. The following products are carrier protein and azide free (catalog numbers 23300002, H00000768-B01P and H00000768-D01P). If you would like to use NB100-417, we do offer antibody purification kits to remove unwanted buffer ingredients. See this link here for those products.
  6. I have a couple of questions regarding the Carbonic Anhydrase IX Antibody (NB100-417) listed on your website. We are wondering if this antibody is carrier-free, and if not, whether you have a carrier-free version?
    • Our NB100-417 is carrier protein free but does contain 0.1% sodium azide. The buffer formulation is: Tris-citrate/phosphate, pH 7-8 with 0.1% sodium azide. The following products are carrier protein and azide free (catalog numbers 23300002, H00000768-B01P and H00000768-D01P). If you would like to use NB100-417, we do offer antibody purification kits to remove unwanted buffer ingredients. See the link here for those products: AbSelect.
  7. I am interested in using the CA9 antibody (NB100-417) that you supply. We dilute our antibodies in 5% milk TBS-Tween and usually freeze-thaw them multiple times. Is this possible with this antibody when diluted?
    • I have checked with the lab and they have informed me that freezing is not recommended for NB100-417 under any circumstances. Unfortunately, the antibody would no longer be covered by our guarantee if it was stored at a temperature that did not match our storage instructions. Of course, you are free to test your proposed conditions if you would like, but we would not be able to accept responsibility if the antibody was damaged by this.
  8. Could this antibody (CA9) be safely stored at -80C?
    • The recommended storage temperature for NB100-417 is -20C. It may be possible to store this antibody at -80C, but this has not been confirmed by our lab. It is most important to ensure that that antibody does not undergo too many freeze-thaw cycles.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB100-417AF405
Alexa Fluor 488 Labeled NB100-417AF488
Alexa Fluor 647 Labeled NB100-417AF647
Alexa Fluor 700 Labeled NB100-417AF700
Biotin Labeled NB100-417B
DyLight 405 Labeled NB100-417V
DyLight 488 Labeled NB100-417G
DyLight 550 Labeled NB100-417R
DyLight 650 Labeled NB100-417C
FITC Labeled NB100-417F
HRP Labeled NB100-417H

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Blogs on Carbonic Anhydrase IX/CA9.

Carbonic anhydrase IX (CAIX) - a reliable histochemical marker of hypoxia
Carbonic anhydrase IX is a member of the carbonic anhydrase family. This family consists of catalytic enzymes capable of converting carbon dioxide and water into carbonic acid, protons, and bicarbonate ions. This family of molecules is abundantly e...  Read full blog post.

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Recent Reviews

4.7
5
2
4
1
3
0
2
0
1
0

 
rajesha rupaimoole
12/10/2014
Application: WB
Species: Human

Xiuquan Luo
07/21/2014
Application: WB
Species: Human

Sonia Rocha
12/21/2011
Application: ICC
Species: Human

Bioinformatics

Gene Symbol CA9
Entrez

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