Antibodies, Proteins, Kits and Reagents for Life Science

307 566 ₸ Product size

100 µl 307 566 ₸ 10 µl 90 460 ₸

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Anti-Pit1 antibody [EPR23555-203] (ab273048)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR23555-203] to Pit1
Suitable for: WB, Flow Cyt, IP, IHC-P, IHC-Fr, ICC/IF
Reacts with: Mouse, Rat

Product name

Anti-Pit1 antibody [EPR23555-203]
See all Pit1 primary antibodies
Description

Rabbit monoclonal [EPR23555-203] to Pit1
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Rat
ICC/IF	Rat
IHC-Fr	Mouse
IHC-P	Mouse
IP	Mouse Rat
WB	Mouse Rat

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: GH3 whole cell lysate; Mouse pituitary tissue. IHC-P: Mouse pituitary tissue. IHC-Fr: Mouse pituitary tissue. ICC/IF: GH3 cells. Flow Cyt: GH3 cells. IP: Mouse pituitary tissue lysate. GH3 whole cell lysate.
General notes
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23555-203
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pit1 antibody [EPR23555-203] (ab273048)

Immunohistochemical analysis of paraffin-embedded mouse pituitary tissue labeling Pit1 with ab273048 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in mouse pituitary (PMID:21123951). Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Frozen sections) - Anti-Pit1 antibody [EPR23555-203] (ab273048)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse pituitary tissue labeling Pit1 with ab273048 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2µg/ml) dilution (Green). Nuclear staining on mouse pituitary is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2µg/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Immunoprecipitation - Anti-Pit1 antibody [EPR23555-203] (ab273048)

Pit1 was immunoprecipitated from 0.35 mg mouse pituitary tissue lysate 10µg with ab273048 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab273048 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Mouse pituitary tissue lysate 10µg.

Lane 2: ab273048 IP in mouse pituitary tissue lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab273048 in mouse pituitary tissue lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 23 seconds.
Western blot - Anti-Pit1 antibody [EPR23555-203] (ab273048)

All lanes : Anti-Pit1 antibody [EPR23555-203] (ab273048) at 1/1000 dilution

Lane 1 : Mouse pituitary tissue lysate
Lane 2 : Mouse liver tissue 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: 33 kDa
Observed band size: 33 kDa

Exposure time: 81 seconds

The expression profile observed is consistent with what has been described in the literature (PMID: 21123951).
Negative control: Mouse liver (PMID: 21123951).

Blocking/Diluting buffer and concentration 5% NFDM/TBST
Western blot - Anti-Pit1 antibody [EPR23555-203] (ab273048)

Anti-Pit1 antibody [EPR23555-203] (ab273048) at 1/1000 dilution + GH3 (Rat pituitary epithelial), whole cell lysate at 20 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 33 kDa
Observed band size: 31,33 kDa
why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

The expression profile observed is consistent with what has been described in the literature. The 31kDa and 33kDa bands correspond to different isoforms (PMID: 21123951, 21060149).

Exposure time: 7 seconds.
Immunocytochemistry/ Immunofluorescence - Anti-Pit1 antibody [EPR23555-203] (ab273048)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized GH3 (Rat pituitary epithelial cell line) cells labelling Pit1 with ab273048 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 2µg/ml dilution (Green). Confocal image showing nuclear staining in GH3 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 2.5µg/ml dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 2µg/ml dilution.
Flow Cytometry - Anti-Pit1 antibody [EPR23555-203] (ab273048)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized GH3 (Rat pituitary epithelial cell line) cells labelling Pit1 with ab273048 at 1/50 dilution (1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-Pit1 antibody [EPR23555-203] (ab273048)

Pit1 was immunoprecipitated from 0.35 mg GH3 (Rat pituitary epithelial), whole cell lysate 10µg with ab273048 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab273048 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: GH3 (Rat pituitary epithelial), whole cell lysate 10µg.

Lane 2: ab273048 IP in GH3 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab273048 in GH3 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 5 seconds.
Immunohistochemistry (Frozen sections) - Anti-Pit1 antibody [EPR23555-203] (ab273048)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver tissue labeling Pit1 with ab273048 at 1/100 dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2µg/ml) dilution (Green). No staining on mouse liver is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 (2µg/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pit1 antibody [EPR23555-203] (ab273048)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling Pit1 with ab273048 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Counterstained with Hematoxylin.

Negative control: No staining in mouse cerebrum (PMID:21123951).

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Anti-Pit1 antibody [EPR23555-203] (ab273048)

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