Antibodies, Proteins, Kits and Reagents for Life Science

Anti-CstF-64 antibody [EPR15698] - BSA and Azide free (ab246326)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR15698] to CstF-64 - BSA and Azide free
Suitable for: WB, IHC-P, IP, ICC/IF, Flow Cyt
Knockout validated
Reacts with: Human

Product name

Anti-CstF-64 antibody [EPR15698] - BSA and Azide free
See all CstF-64 primary antibodies
Description

Rabbit monoclonal [EPR15698] to CstF-64 - BSA and Azide free
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, IP, ICC/IF, Flow Cytmore details
Species reactivity

Reacts with: Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- Human testis tissue lysate.IHC: Human cervix carcinoma and Human tonsil tissue.ICC/IF: HeLa and Jurkat cells.IP: K562 whole cell lysate.
General notes
Ab246326 is the carrier-free version of ab200837. 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
ab246326 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

EPR15698
Isotype

IgG
Research areas

Western blot - Anti-CstF-64 antibody [EPR15698] - BSA and Azide free (ab246326)

All lanes : Anti-CstF-64 antibody [EPR15698] (ab200837) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CSTF2 knockout HeLa cell lysate
Lane 3 : K-562 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 61 kDa
Observed band size: 65 kDa
why is the actual band size different from the predicted?

This data was developed using the same antibody clone in a different buffer formulation (ab200837).

Lanes 1-3: Merged signal (red and green). Green - ab200837 observed at 65 kDa. Red - loading control ab8245 observed at 36 kDa.

ab200837 Anti-CstF-64 antibody [EPR15698] was shown to specifically react with CstF-64 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265207 (knockout cell lysate ab257402) was used. Wild-type and CstF-64 knockout samples were subjected to SDS-PAGE. ab200837 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CstF-64 antibody [EPR15698] - BSA and Azide free (ab246326)

Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling CstF-64 with ab200837 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on Human cervix carcinoma 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200837).

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-CstF-64 antibody [EPR15698] - BSA and Azide free (ab246326)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CstF-64 with ab200837 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on Human tonsil tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200837).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-CstF-64 antibody [EPR15698] - BSA and Azide free (ab246326)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling CstF-64 with ab200837 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear 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: ab200837 at 1/500 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200837).
Immunocytochemistry/ Immunofluorescence - Anti-CstF-64 antibody [EPR15698] - BSA and Azide free (ab246326)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling CstF-64 with ab200837 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear 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: ab200837 at 1/500 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200837).
Immunoprecipitation - Anti-CstF-64 antibody [EPR15698] - BSA and Azide free (ab246326)

CstF-64 was immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate with ab200837 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab200837 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: K562 whole cell lysate10 µg (Input). Lane 2: ab200837 IP in K562 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200837 in K562 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 5 seconds

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200837).
Flow Cytometry - Anti-CstF-64 antibody [EPR15698] - BSA and Azide free (ab246326)

Flow Cytometry analysis of Jurkat (human acute T cell leukemia) labelling CstF-64 with purified ab200837 at 1/25000 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Alexa Fluor® 488 goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab200837).
Anti-CstF-64 antibody [EPR15698] - BSA and Azide free (ab246326)

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