Recombinant Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562)

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Western blot - Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR27167-58] to Calcineurin B/CNB + PPP3R2
Suitable for: IHC-P, WB, IP
Reacts with: Mouse, Rat, Human

Product name

Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14]
Description

Rabbit monoclonal [EPR27167-58] to Calcineurin B/CNB + PPP3R2
Host species

Rabbit
Tested applications

Suitable for: IHC-P, WB, IPmore details
Unsuitable for: Flow Cyt (Intra) or ICC/IF
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human testis, cerebellum, kidney, and heart tissue lysates; mouse and rat testis, brain, kidney, and heart tissue lysates; HeLa, C6, PC12 whole cell lysates; HEK-293T cells transfected with a human (Calcineurin B/PPP3R1 or Calcineurin BII/PPP3R2) expression vector containing a his tag, whole cell lysate. IHC-P: Human, mouse, and rat cerebrum FFPE tissue sections. IP: HEL (human erythroleukemia erythroblast), 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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR27167-58
Isotype

IgG
Research areas

Alternative Versions
- Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] - BSA and Azide free (ab303563)
Compatible Secondaries
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab303562 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
IHC-P		1/10000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB		1/1000. Detects a band of approximately 15, 18, 20 kDa (predicted molecular weight: 19 kDa).
IP		1/30.

Notes
IHC-P 1/10000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 15, 18, 20 kDa (predicted molecular weight: 19 kDa).
IP 1/30.

Application notes

Is unsuitable for Flow Cyt (Intra) or ICC/IF.

Western blot - Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562)

All lanes : Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562) at 1/1000 dilution

Lane 1 : HEK-293T cells transfected with a human Calcineurin B/PPP3R1 expression vector containing a his tag, whole cell lysate at 1 µg
Lane 2 : HEK-293T cells transfected with a human Calcineurin BII/PPP3R2 expression vector containing a his tag, whole cell lysate at 75 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 19 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?

Blocking / Diluting buffer and concentration: 5% NFDM/TBST

The identity of the higher MW band at approximately 75kDa is unknown.

Exposure: 7.75 seconds.
Western blot - Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562)

All lanes : Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562) at 1/1000 dilution

Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse kidney tissue lysate
Lane 3 : Mouse heart tissue lysate
Lane 4 : Rat brain tissue lysate
Lane 5 : Rat kidney tissue lysate
Lane 6 : Rat heart tissue lysate

Lysates/proteins at 40 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 19 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted?

Exposure time: 10 seconds

Blocking / Diluting buffer and concentration: 5% NFDM/TBST

Low expression: heart, kidney (PMID:8110831).

The identity of the higher MW band at approximately 75 kDa is unknown.
Western blot - Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562)

All lanes : Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562) at 1/1000 dilution

Lane 1 : Human testis tissue lysate
Lane 2 : Mouse testis tissue lysate
Lane 3 : Rat testis tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 19 kDa
Observed band size: 15,18 kDa why is the actual band size different from the predicted?

Exposure time: 136 seconds

Blocking / Diluting buffer and concentration: 5% NFDM/TBST

The higher MW band at approximately 18 kDa (in lane 2) is PPP3R2. (PMID: 26429887)
Western blot - Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562)

All lanes : Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562) at 1/1000 dilution

Lane 1 : Human cerebellum tissue lysate
Lane 2 : Human kidney tissue lysate
Lane 3 : Human heart tissue lysate

Lysates/proteins at 40 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 19 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted?

Blocking / Diluting buffer and concentration: 5% NFDM/TBST

Low expression: heart, kidney (PMID:8110831).

The molecular weight observed is consistent with what has been described in the literature (PMID: 30262862).

The identity of the higher MW band at approximately 75 kDa (in lane 3) is unknown.
Western blot - Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562)

All lanes : Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562) at 1/1000 dilution

Lane 1 : HEL (human erythroleukemia erythroblast), whole cell lysate
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate

Lysates/proteins at 40 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 19 kDa
Observed band size: 15 kDa why is the actual band size different from the predicted?

Exposure time: 70 seconds

Blocking / Diluting buffer and concentration: 5% NFDM/TBST
Immunoprecipitation - Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562)

Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 was immunoprecipitated from 0.35 mg HEL (human erythroleukemia erythroblast), whole cell lysate with ab303562 at 1/30 dilution (2 µg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab303562 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP) (ab131366) was used at 1/5000 dilution.

Lane 1: HEL (human erythroleukemia erythroblast), whole cell lysate (Inset)

Lane 2: ab303562 IP in HEL whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of 303562 in HEL whole cell lysate

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

Exposure time: 110 seconds

This blot was developed using a high sensitivity ECL substrate.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 with ab303562 at 1/10000 dilution (0.052 µg/mL), followed by a ready to use LeicaDS9800 dilution (BOND^™ Polymer Refine Detection). Negative control: no staining is observed on human kidney. The section was incubated with ab303562 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 dilution (BOND^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562)

Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 with ab303562 at 1/10000 dilution (0.052 µg/mL), followed by a ready to use LeicaDS9800 dilution (BOND^™ Polymer Refine Detection). Positive staining was observed on rat cerebrum. The section was incubated with ab303562 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 dilution (BOND^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 with ab303562 at 1/10000 dilution (0.052 µg/mL), followed by a ready to use LeicaDS9800 dilution (BOND^™ Polymer Refine Detection). Positive staining was observed on mouse cerebrum. The section was incubated with ab303562 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 dilution (BOND^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 with ab303562 at 1/10000 dilution (0.052 µg/mL), followed by a ready to use LeicaDS9800 dilution (BOND^™ Polymer Refine Detection). Positive staining was observed on human cerebrum. The section was incubated with ab303562 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 dilution (BOND^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins
Anti-Calcineurin B/PPP3R1 + Calcineurin BII/PPP3R2 antibody [EPR24992-14] (ab303562)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

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Datasheet download

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Publishing research using ab303562? Please let us know so that we can cite the reference in this datasheet.

ab303562 has not yet been referenced specifically in any publications.

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Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Applications

The Abpromise guarantee

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (0)

Customer reviews and Q&As