Antibodies, Proteins, Kits and Reagents for Life Science

Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19803] to HP1 gamma/CBX3 - BSA and Azide free
Suitable for: Flow Cyt, IHC-P, WB, ICC
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free
See all HP1 gamma/CBX3 primary antibodies
Description

Rabbit monoclonal [EPR19803] to HP1 gamma/CBX3 - BSA and Azide free
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
IHC-P	Mouse
WB	Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
General notes
ab251540 is the carrier-free version of ab213167 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
Ab251540 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Maxpar® is a trademark of Fluidigm Canada Inc.

This product was previously labelled as HP1 gamma
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...
Clonality

Monoclonal
Clone number

EPR19803
Isotype

IgG
Research areas

Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)

This data was developed using ab213167, the same antibody clone in a different buffer formulation.

Lane 1: Wild type HAP1 whole cell lysate (20 µg)

Lane 2: CBX3 (KO) knockout HAP1 whole cell lysate (20 µg)

Lane 3: HUVEC whole cell lysate (20 µg)

Lane 4: HepG2 whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab213167 observed at 23 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab213167 was shown to recognize HP1 gamma/CBX3 when HP1 gamma/CBX3 knockout samples were used, along with additional cross-reactive bands. Wild-type and HP1 gamma/CBX3 knockout samples were subjected to SDS-PAGE. ab213167 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 500 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)

All lanes : Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167) at 1/1000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 21 kDa
Observed band size: 21 kDa

Exposure time: 3 minutes

This data was developed using ab213167, the same antibody clone in a different buffer formulation.
Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)

All lanes : Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167) at 1/1000 dilution

Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 21 kDa
Observed band size: 21 kDa

This data was developed using ab213167, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure times: Lane 1: 3 minutes; Lane 2: 15 seconds.
Western blot - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)

All lanes : Anti-HP1 gamma/CBX3 antibody [EPR19803] (ab213167) at 1/1000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Mouse spleen lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat spleen lysate
Lane 6 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 7 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 8 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 9 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 21 kDa
Observed band size: 21 kDa

This data was developed using ab213167, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: Lanes 1 and 2: 3 minutes; Lane 3: 10 seconds; Lane 4: 30 seconds; Lane 5: 10 seconds; Lanes 6-9: 30 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)

This data was developed using ab213167, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling HP1 gamma/CBX3 with ab213167 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on epithelial cells of Human colon is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution. Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)

This data was developed using ab213167, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling HP1 gamma/CBX3 with ab213167 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on cancer cells of Human cervix carcinoma is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution. Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)

This data was developed using ab213167, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling HP1 gamma/CBX3 with ab213167 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on Mouse testis is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution. Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Immunocytochemistry - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)

This data was developed using ab213167, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling HP1 gamma/CBX3 with ab213167 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on NIH/3T3 cells. Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red). The negative controls are as follows:- -ve control 1: ab213167 at 1/100 dilution followed by ab150120 at 1/1000 dilution. -ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
Immunocytochemistry - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)

This data was developed using ab213167, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling HP1 gamma/CBX3 with ab213167 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cells. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red). The negative controls are as follows:- -ve control 1: ab213167 at 1/100 dilution followed by ab150120 at 1/1000 dilution. -ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
Flow Cytometry - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)

This data was developed using ab213167, the same antibody clone in a different buffer formulation.Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling HP1 gamma/CBX3 with ab213167 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal-Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.
Flow Cytometry - Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)

This data was developed using ab213167, the same antibody clone in a different buffer formulation.Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling HP1 gamma/CBX3 with ab213167 at 1/60 dilution (red) compared with a Rabbit IgG,monoclonal - Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.
Anti-HP1 gamma/CBX3 antibody [EPR19803] - BSA and Azide free (ab251540)

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