Antibodies, Proteins, Kits and Reagents for Life Science

Anti-Alpha B Crystallin antibody [EPR24521-60] (ab281561)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR24521-60] to Alpha B Crystallin
Suitable for: Flow Cyt (Intra), ICC, IHC-P, IP, WB
Reacts with: Mouse, Rat, Human

Product name

Anti-Alpha B Crystallin antibody [EPR24521-60]
See all Alpha B Crystallin primary antibodies
Description

Rabbit monoclonal [EPR24521-60] to Alpha B Crystallin
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), ICC, IHC-P, IP, WBmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human heart, MDA-MB-468, Rat heart, Mouse heart, brain, lung, HL-1, C2C12 lysates. IHC-P: Human breast, Human cardiac muscle, Mouse cerebrum, cardiac muscle, Rat kidney tissues. ICC: HL-1, C2C12 cells. Flow cyt: HL-1 cell. IP: Mouse heart 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.
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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR24521-60
Isotype

IgG

Western blot - Anti-Alpha B Crystallin antibody [EPR24521-60] (ab281561)

All lanes : Anti-Alpha B Crystallin antibody [EPR24521-60] (ab281561) at 1/5000 dilution

Lane 1 : Human heart tissue lysate
Lane 2 : MDA-MB-468 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : Rat heart tissue lysate

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 20 kDa
Observed band size: 21 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 (PMID: 17661394, 15197794, 24710637).

Negative control: MCF7 (PMID:15197794, 24710637).

Exposure time: Lane 1:92 seconds; Lanes 2-3: 3 minutes; Lane 4: 3 seconds.
Western blot - Anti-Alpha B Crystallin antibody [EPR24521-60] (ab281561)

All lanes : Anti-Alpha B Crystallin antibody [EPR24521-60] (ab281561) at 1/1000 dilution

Lane 1 : Mouse heart tissue lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Mouse lung tissue lysate
Lane 4 : HL-1 (mouse atrial muscle cell) whole cell lysate
Lane 5 : C2C12 (mouse myoblasts myoblast) 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 (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 20 kDa
Observed band size: 21 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 (PMID: 17661394, 15197794, 24710637).

Low expression: Mouse lung (PMID:17661394).

Exposure time: Lane 1:26 seconds; Lanes 2-4: 3 minutes; Lane 5: 26 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha B Crystallin antibody [EPR24521-60] (ab281561)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labelling Alpha B Crystallin with ab281561 at 1/3000 dilution (0.089 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human breast tissues (PMID:24987308). The section was incubated with ab281561 for 10 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha B Crystallin antibody [EPR24521-60] (ab281561)

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labelling Alpha B Crystallin with ab281561 at 1/3000 dilution (0.089 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human cardiac muscle. The section was incubated with ab281561 for 10 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha B Crystallin antibody [EPR24521-60] (ab281561)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling Alpha B Crystallin with ab281561 at 1/5000 dilution (0.089 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cerebrum (PMID:1739128). The section was incubated with ab281561 for 10 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha B Crystallin antibody [EPR24521-60] (ab281561)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labelling Alpha B Crystallin with ab281561 at 1/3000 dilution (0.089 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cardiac muscles (PMID:27566162). The section was incubated with ab281561 for 10 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha B Crystallin antibody [EPR24521-60] (ab281561)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labelling Alpha B Crystallin with ab281561 at 1/3000 dilution (0.089 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat kidney (PMID:14566023). The section was incubated with ab281561 for 10 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunocytochemistry - Anti-Alpha B Crystallin antibody [EPR24521-60] (ab281561)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HL-1 (mouse atrial muscle cell) cells labelling Alpha B Crystallin with ab281561 at 1/50 dilution (8.88 ug/ml), followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HL-1 cells is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 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 dilution.
Immunocytochemistry - Anti-Alpha B Crystallin antibody [EPR24521-60] (ab281561)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C2C12 (mouse myoblasts myoblast) cells labelling Alpha B Crystallin with ab281561 at 1/50 dilution (8.88 ug/ml), followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in C2C12 cells is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 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 dilution.
Flow Cytometry (Intracellular) - Anti-Alpha B Crystallin antibody [EPR24521-60] (ab281561)

Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HL-1 (mouse atrial muscle cell) cells labelling Alpha B Crystallin with ab281561 at 1/500 dilution (0.1ug)(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). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-Alpha B Crystallin antibody [EPR24521-60] (ab281561)

Alpha B Crystallin was immunoprecipitated from 0.35 mg Mouse heart tissue lysate with ab281561 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab281561 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Mouse heart tissue lysate 10 ug

Lane 2: ab281561 IP in Mouse heart tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab281561 in mouse heart tissue lysate

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

Exposure time: 6 seconds.
Anti-Alpha B Crystallin antibody [EPR24521-60] (ab281561)

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