Name: Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker
SKU: ab229025
Rating: 4 (5 reviews)

Rabbit Recombinant Monoclonal Lamin B1 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), mIHC, IHC-P and reacts with Mouse, Human, Rat samples. Cited in 12 publications.

Images

Western blot - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (AB229025), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Abcam Recommends

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	Flow Cyt (Intra)	mIHC	IHC-P
Human	Tested	Tested	Tested	Tested	Tested	Tested
Mouse	Tested	Tested	Tested	Tested	Expected	Tested
Rat	Expected	Tested	Expected	Expected	Expected	Tested

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -
Species Human	Dilution info 1/500	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/4000	Notes Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/2000	Notes Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/2000	Notes Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/2000	Notes Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

Select an associated product type

11 products for Alternative Product

Target data

See full target information LMNB1

Function

Lamins are intermediate filament proteins that assemble into a filamentous meshwork, and which constitute the major components of the nuclear lamina, a fibrous layer on the nucleoplasmic side of the inner nuclear membrane (PubMed:28716252, PubMed:32910914). Lamins provide a framework for the nuclear envelope, bridging the nuclear envelope and chromatin, thereby playing an important role in nuclear assembly, chromatin organization, nuclear membrane and telomere dynamics (PubMed:28716252, PubMed:32910914). The structural integrity of the lamina is strictly controlled by the cell cycle, as seen by the disintegration and formation of the nuclear envelope in prophase and telophase, respectively (PubMed:28716252, PubMed:32910914).

Alternative names

LMN2, LMNB, LMNB1, Lamin-B1

Recommended products

Rabbit Recombinant Monoclonal Lamin B1 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), mIHC, IHC-P and reacts with Mouse, Human, Rat samples. Cited in 12 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR22165-121
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Lamin B1 also known as LMNB1 is a protein that plays an important role in the structure of the nuclear lamina a dense fibrillar network inside the nucleus of cells. This protein belongs to the lamin family and has an approximate molecular weight of 66 kDa. Lamin B1 is expressed in nearly all types of cells acting as a structural component by interacting with chromatin and anchoring the nuclear envelope. It contributes to regulating nuclear stability and mechanical support which is essential for proper cell function.

Biological function summary

Lamin B1 serves many important functions by providing structural integrity and regulating gene expression through chromatin organization. Part of the nuclear lamina complex Lamin B1 interacts with other lamin proteins such as Lamin A and Lamin C forming a mesh-like structure that supports the nuclear envelope. This protein also affects cellular processes such as DNA replication RNA transcription and cell division indicating its broad impact on various cellular activities.

Pathways

Lamin B1 plays an important role in gene expression regulation and cell cycle control. It interacts with pathways involved in chromatin remodeling influencing the access of transcription factors to DNA which affects gene expression profiles. Lamin B1 associates with related proteins such as Lamin A/C and other nuclear matrix proteins to maintain cellular architecture stability and normal cell function. This organization is vital to ensure the accuracy of cellular processes and prevent anomalies during cell division.

Associated diseases and disorders

Mutations or altered expression of Lamin B1 have been associated with several pathological conditions. One noteworthy disorder is autosomal dominant adult-onset leukodystrophy where Lamin B1 expression levels are abnormal. This protein also connects to multiple sclerosis through its role in maintaining nuclear architecture. Additionally changes in Lamin B1 expression levels can influence pathways involving Lamin A/C and potentially lead to other laminopathies highlighting its influence in maintaining cellular health and contributing to disease development.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

16 product images

Western blot - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025)
Lamin B1 Western blot staining using rabbit Anti-Lamin B1 antibody
Lanes 1- 2: Merged signal (red and green). Green - ab229025 observed at 66-70 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
ab229025 was shown to react with Lamin B1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human LMNB1 (Lamin B1) knockout HeLa cell line ab255404 (knockout cell lysate Human LMNB1 (Lamin B1) knockout HeLa cell lysate ab263825) was used. Wild-type HeLa and LMNB1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab229025 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (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.
All lanes: Western blot - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: LMNB1 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human LMNB1 (Lamin B1) knockout HeLa cell line (Human LMNB1 (Lamin B1) knockout HeLa cell line ab255404)
Performed under reducing conditions.
Predicted band size: 66 kDa
Observed band size: 66-70 kDa
Western blot - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025)
Lamin B1 Western blot staining using rabbit Anti-Lamin B1 antibody
ab229025 was shown to specifically react with Lamin B1 in wild-type HAP1 cells as signal was lost in Lamin B1 knockout cells. Wild-type and Lamin B1 knockout samples were subjected to SDS-PAGE. ab229025 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD^® ChemiDoc™ MP instrument using the ECL technique.
All lanes: Western blot - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025) at 1/5000 dilution
Lane 1: Wild-type HAP1 whole cell lysate at 20 µg
Lane 2: Lamin B1 knockout HAP1 whole cell lysate at 20 µg
Lane 3: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 4: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg
Predicted band size: 66 kDa
Observed band size: 70 kDa
Exposure time: 8s
Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025)
Lamin B1 Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-Lamin B1 antibody
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Lamin B1 (Green) with ab229025 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution. Confocal image showing nuclear membranous staining in HeLa cells. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)(Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) was used as a counterstain at 1/200 dilution. The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Alexa Fluor^®488 Goat anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody [EPR22165-121] (ab229025)
Immunohistochemical analysis of paraffin-embedded mouse liver tissue stained for Lamin B1 using ab229025 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear envelope staining on mouse liver (PMID: 19925772) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025)
Lamin B1 Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-Lamin B1 antibody
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast cell line) cells labeling Lamin B1 (Green) with ab229025 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution. Confocal image showing nuclear membranous staining in NIH/3T3 cells. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)(Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) was used as a counterstain at 1/200 dilution. The nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Alexa Fluor^®488 Goat anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.
Immunoprecipitation - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025)
Lamin B1 was immunoprecipitated from 0.35 mg HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab229025 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab229025 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/5000 dilution.
Lane 1: HeLa whole cell lysate 10 μg (input).
Lane 2: ab229025 IP in HeLa whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab229025 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
A very weak band around 66 kDa detected in lane 3 is due to spill over from lane2 (+).
All lanes: Immunoprecipitation - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025)
Predicted band size: 66 kDa
Observed band size: 66 kDa
Multiplex immunohistochemistry - Anti-Lamin B1 antibody [EPR22165-121] (ab229025)
Multiplex immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue.
Panel A: Merged staining of Collagen VI (Anti-Collagen VI antibody [EPR17072] ab182744; green), anti-CD68 (Anti-CD68 antibody [EPR20545] ab213363; red) and anti-Lamin B1 (ab229025; magenta).
Panel B: Anti-Collagen VI (green) stained on extracellular matrix.
Panel C: Anti-CD68 (red) stained on Kupffer cells.
Panel D: Anti-Lamin B1 (magenta) stained on nuclear envelope.
Key protocol steps: The section was incubated in three rounds of staining with Anti-Collagen VI antibody [EPR17072] ab182744 (1/1000 dilution), Anti-CD68 antibody [EPR20545] ab213363 (1/1000 dilution) and ab229025 (1/4000 dilution) for 30 mins at room temperature. Each round was followed by tyramide signal amplification with the appropriate fluorophore. Heat mediated antigen retrieval was used (Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins after every round of antibody/fluorophore staining.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
DAPI was used as a nuclear counter stain. A ready-to-use anti-Rabbit and Mouse Polymer HRP was used as a secondary.
Western blot - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025)
Lamin B1 Western blot staining using rabbit Anti-Lamin B1 antibody
Blocking/diluting buffer and concentration: 5% NFDM/TBST.
Exposure time:
Lane 1:3 seconds;
Lanes 2,3,4 and 5:10 seconds.
All lanes: Western blot - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025) at 1/1000 dilution
Lane 1: Human lymph node tissue lysate at 20 µg
Lane 2: HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 3: HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Lane 4: Caco-2 (Human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 5: Mouse brain tissue lysate at 20 µg
Secondary
Lane 1: Western blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/100000 dilution
Lanes 2 - 5: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 66 kDa
Observed band size: 70 kDa
Immunoprecipitation - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025)
Lamin B1 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate with ab229025 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab229025 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/5000 dilution.
Lane 1: NIH/3T3 whole cell lysate 10 μg (input).
Lane 2: ab229025 IP in NIH/3T3 whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab229025 in NIH/3T3 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
All lanes: Immunoprecipitation - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025)
Predicted band size: 66 kDa
Observed band size: 66 kDa
Western blot - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025)
Lamin B1 Western blot staining using rabbit Anti-Lamin B1 antibody
Blocking/diluting buffer and concentration: 5% NFDM/TBST.
Exposure time:
Lane 1:10 seconds; Lanes 2 and 3: 59 seconds; Lane 4:3 seconds; Lanes 5,6 and 7:6 seconds.
All lanes: Western blot - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025) at 1/1000 dilution
Lane 1: Mouse heart tissue lysate at 10 µg
Lane 2: Rat brain tissue lysate at 10 µg
Lane 3: Rat heart tissue lysate at 10 µg
Lane 4: Rat spleen tissue lysate at 10 µg
Lane 5: RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg
Lane 6: PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg
Lane 7: NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 66 kDa
Observed band size: 70 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025)
Lamin B1 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-Lamin B1 antibody
Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue stained for Lamin B1 using ab229025 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear envelope staining on rat cerebrum (PMID: 26469707, PMID: 19925772) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody [EPR22165-121] (ab229025)
Immunohistochemical analysis of paraffin-embedded human breast tissue stained for Lamin B1 using ab229025 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear envelope staining on human breast (PMID: 26469707) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Flow Cytometry (Intracellular) - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025)
Lamin B1 Flow Cytometry (Intracellular) staining using rabbit Anti-Lamin B1 antibody
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling Lamin B1 with ab229025 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgGisotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^ï¿½ 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/2000 dilution.
Western blot - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025)
Lamin B1 Western blot staining using rabbit Anti-Lamin B1 antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
In Western blot, Anti-Lamin B1 antibody [EPR22165-121] (ab229025) staining at 1/1000 dilution.
All lanes: Western blot - Anti-Histone H3 antibody [RM1288] - Nuclear Marker (Anti-Histone H3 antibody [RM1288] - Nuclear Marker ab322707) at 1/10000 dilution
Lane 1: HEK-293 (human embryonic kidney epithelial cell) nuclear fraction at 10 µg
Lane 2: HEK-293 cytoplasmic fraction at 10 µg
Lane 3: NIH/3T3 (mouse embryonic fibroblast) nuclear fraction at 10 µg
Lane 4: NIH/3T3 cytoplasmic fraction at 10 µg
Lane 5: Rat brain tissue nuclear fraction at 10 µg
Lane 6: Rat brain tissue cytoplasmic fraction at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 16 kDa, 14 kDa, 36 kDa, 66 kDa
Exposure time: 8s
Flow Cytometry (Intracellular) - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025)
Lamin B1 Flow Cytometry (Intracellular) staining using rabbit Anti-Lamin B1 antibody
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cell line labeling Lamin B1 with ab229025 at 1/500 dilution (Red) compared with aRabbit monoclonal IgGisotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^ï¿½ 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/2000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody [EPR22165-121] - Nuclear Envelope Marker (ab229025)
Lamin B1 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-Lamin B1 antibody
Immunohistochemical analysis of paraffin-embedded human endometrial carcinoma stained for Lamin B1 using ab229025 at 1/2000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Nuclear envelope staining on human endometrial carcinoma is observed. Counterstained with hematoxylin.

Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). The section was incubated with ab229025 at 4°C overnight