Can anyone suggest a good Lamin B1 antibody that actually works well in IF microscopy?

Dear Ilze!

You can definitely find such antibody using Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker

It is the Recombinant RabMAbKO

Validated. 20ul selling size

- Cited in over 185 publications - Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests - Specificity confirmed with LMNB1 knockout cell line validation - Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation - Antibody clone EPR8985(B) is cited in over 250 publications - Affinity purified using Protein AAnti-lamin B1 antibody [EPR8985(B)] is a rabbit monoclonal antibody that is used in lamin B1 western blot (WB), IHC, immunoprecipitation (IP) and immunocytochemistry/immunofluorescence (ICC/IF). Suitable for human, mouse and rat samples.

All the best!

Ilya B. Tsyrlov, MD, PhD

President, Xenotox, Inc.

https://xenobiovir.com/

Lucian Miles

Hi Ilze, Lamin B1 Antibody (MCE, HY-P80205) is suitable for IF-Tissue. Lamin B1 Antibody is a non-conjugated and Rabbit origined monoclonal antibody about 66 kDa, targeting to Lamin B1. It can be used for WB、IHC-P、IF-Tissue assays with tag free, in the background of Human, Mouse, Rat.

Hope this helpful and feel free to reach out.

https://www.medchemexpress.com/antibody/lamin-b1-rabbit-mab.html

Ilze Skujina

Lucian Miles thank you.

Would you know the distributer in Ireland?

And would you mind providing a short protocol (ie ethanol or PFA fixation, incubation time and temp please)?

I have tried 2 Lamin B1 antibodies from Abcam and Cell Signalling that are supposed to work for IF, but they work only for Westerns, not IF unfortunately and my colleagues also are struggling to find one that works well in IF.

Lucian Miles

Dear Ilze,

Thank you for reaching out to us with your question. We’d be delighted to assist you! Regarding your IF experimental protocol for the Lamin B1 antibody and your request for distributor information in Ireland, I’ve relayed these details to our team. Since some technical specifics are involved, would you kindly share your email address with us—either in the comments or via private message? Our dedicated team will promptly follow up with you via email.

Alternatively, you’re welcome to contact us directly at [email protected].

Please don’t hesitate to reach out if you have any further questions. We’re here to help!

Lucian Miles

Please find the IF protocol and experimental conditions for your reference. For the Lamin B1 Antibody (MCE, HY-P80205), we recommend trying EDTA-based heat-induced antigen retrieval with boiling method, using a primary antibody dilution ratio of 1:100.

For more details about the MCE Lamin B1 Antibody or customized experimental solutions, please feel free to discuss with us via emails.

Immunofluorescence (IF) Staining Protocol

I. Paraffin Section Pretreatment

Dewaxing: Immerse slides sequentially in: Xylene I (15 min) → Xylene II (15 min) Absolute ethanol I (3 min) → Absolute ethanol II (3 min) 95% ethanol (3 min)

Rehydration: Rinse under running tap water for 3 min to complete rehydration.

II. Antigen Retrieval

(Note: Retrieval method should be optimized based on target antigen.)

Option 1: Microwave Retrieval (Tris-EDTA Buffer, pH 9.0)

Fill a beaker with ~650 mL of 1× Tris-EDTA buffer (pH 9.0; adjust volume for slide quantity) and heat in a microwave at high power until boiling.

Submerge slides in preheated buffer using a retrieval rack.

Maintain at 95–100°C (low power) for 20 min (split into 10-min intervals). Ensure slides remain fully immersed; remove bubbles if needed.

Cool at room temperature (RT) for 10 min, then rinse under tap water until RT is reached.

Option 2: Heat-Induced Epitope Retrieval (HIER; Citrate Buffer, pH 6.0)

Add 1 L of 1× citrate buffer (pH 6.0) to a pressure cooker and heat (unsealed) until boiling.

Load slides, seal the valve, and heat until steam releases steadily.

Maintain full pressure for 2 min. Cool gradually by spraying water until the valve drops.

Open the lid and rinse slides under tap water to RT.

III. Blocking

Lightly dry slides and circle tissues with a hydrophobic barrier pen to confine reagents.

Apply 10% normal goat serum (NGS) to fully cover tissues.

Incubate horizontally in a humidified chamber at RT for 1 hr.

IV. Primary Antibody Incubation

Tap off excess serum and apply 100 μL primary antibody (diluted in 10% NGS at optimized concentration). Negative control: Replace primary antibody with PBS.

Incubate in a humidified chamber at 4°C overnight (or 1 hr at RT for validated protocols).

V. Secondary Antibody Incubation

Wash slides 3× with PBS (pH 7.4), 3 min per wash.

Tap dry, then apply 100 μL fluorescently labeled secondary antibody (e.g., goat anti-mouse/rabbit IgG) containing nuclear stain (1:2000 DAPI or Hoechst 33258).

Incubate at RT for 1 hr (protected from light).

VI. Mounting

Wash slides 3× with PBS (pH 7.4), 3 min per wash.

Mount with PBS (pH 7.4) or anti-fade mounting medium. PBS (pH 7.4), or Anti-Fade Mounting Medium.

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