Antibodies, Proteins, Kits and Reagents for Life Science

Product name

Anti-NCAPH2 antibody [EPR17170] - BSA and Azide free
See all NCAPH2 primary antibodies
Description

Rabbit monoclonal [EPR17170] to NCAPH2 - BSA and Azide free
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
IHC-P	Human
IP	Human
WB	Mouse

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
General notes
Ab251316 is the carrier-free version of ab200659. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
ab251316 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C. Do Not Freeze.
Storage buffer

pH: 7.2
Constituent: PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR17170
Isotype

IgG
Research areas

Immunocytochemistry - Anti-NCAPH2 antibody [EPR17170] - BSA and Azide free (ab251316)

This data was developed using ab200659, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling NCAPH2 with ab200659 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).Nuclear and weakly cytoplasm staining on HeLa cell line is observed.The nuclear counter stain is DAPI (blue).Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab200659 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Western blot - Anti-NCAPH2 antibody [EPR17170] - BSA and Azide free (ab251316)

Anti-NCAPH2 antibody [EPR17170] (ab200659) at 1/10000 dilution + Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 68 kDa
Observed band size: 68 kDa

Exposure time: 15 seconds

This data was developed using ab200659, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.
Flow Cytometry - Anti-NCAPH2 antibody [EPR17170] - BSA and Azide free (ab251316)

This data was developed using ab200659, the same antibody clone in a different buffer formulation.
ab200659 staining NCAPH2 in Jurkat (human acute T cell leukemia) cells by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/240. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody. Isoytype control: Rabbit monoclonal IgG (Black) Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
Western blot - Anti-NCAPH2 antibody [EPR17170] - BSA and Azide free (ab251316)

All lanes : Anti-NCAPH2 antibody [EPR17170] (ab200659) at 1/1000 dilution

Lane 1 : Mouse heart lysate
Lane 2 : Rat heart lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 68 kDa
Observed band size: 68 kDa

Exposure time: 3 minutes

This data was developed using ab200659, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-NCAPH2 antibody [EPR17170] - BSA and Azide free (ab251316)

All lanes : Anti-NCAPH2 antibody [EPR17170] (ab200659) at 1/1000 dilution

Lane 1 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 68 kDa
Observed band size: 68 kDa

Exposure time: 15 seconds

This data was developed using ab200659, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAPH2 antibody [EPR17170] - BSA and Azide free (ab251316)

This data was developed using ab200659, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling NCAPH2 with ab200659 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and weakly cytoplasm staining on Human tonsil tissue is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAPH2 antibody [EPR17170] - BSA and Azide free (ab251316)

This data was developed using ab200659, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human clear cell carcinoma of kidney tissue labeling NCAPH2 with ab200659 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and cytoplasm staining on Human clear cell carcinoma of kidney tissue is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAPH2 antibody [EPR17170] - BSA and Azide free (ab251316)

This data was developed using ab200659, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling NCAPH2 with ab200659 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and weakly cytoplasmic staining on Human hepatocellular carcinoma is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCAPH2 antibody [EPR17170] - BSA and Azide free (ab251316)

This data was developed using ab200659, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling NCAPH2 with ab200659 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and weakly cytoplasmic staining on rat kidney is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry - Anti-NCAPH2 antibody [EPR17170] - BSA and Azide free (ab251316)

This data was developed using ab200659, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling NCAPH2 with ab200659 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).Nuclear and weakly cytoplasm staining on Jurkat cell line is observed.The nuclear counter stain is DAPI (blue).Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab200659 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Immunoprecipitation - Anti-NCAPH2 antibody [EPR17170] - BSA and Azide free (ab251316)

This data was developed using ab200659, the same antibody clone in a different buffer formulation.NCAPH2 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab200659 at 1/60 dilution.Western blot was performed from the immunoprecipitate using ab200659 at 1/1000 dilution.Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.Lane 1: HeLa whole cell lysate 10 µg (Input).Lane 2: ab200659 IP in HeLa whole cell lysate.Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200659 in HeLa whole cell extract.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Anti-NCAPH2 antibody [EPR17170] - BSA and Azide free (ab251316)