Thermo Fisher Western Blot: PVDF/NC Membranes, Protein Ladders & ECL Reagents

As a Thermo Fisher distributor, Iright brings together everything you need to run consistent, publish-ready western blots—from membranes and transfer stacks to ECL substrates, ladders, buffers, and stripping solutions. This collection page helps you compare options quickly and route to the right SKUs for your workflow and budget.

Build a Reliable Western Blot Workflow

Western blot quality is decided step by step: clean sample prep, crisp separation, efficient transfer, and matched detection. Below is a practical, four-stage view so you can diagnose issues and choose the right consumables without guesswork.

• Prepare → Separate (SDS-PAGE): Confirm protein load and buffer compatibility; use a broad-range ladder for band tracking.
• Transfer to PVDF/NC: Pick pore size based on protein MW (0.2 µm for small targets; 0.45 µm for mid–high MW); ensure consistent contact and cooling.
• Block → Probe: Select blocking chemistry (BSA/casein/commercial) aligned to HRP chemiluminescence or fluorescent detection; validate host/target for primaries and conjugates for secondaries.
• Detect → Quantify: Choose ECL sensitivity (regular → femto) or NIR fluorescence; keep exposures within the linear range; document settings for reproducibility.

Core Product Categories & How to Choose

Every category below maps to a decision you make at the bench: binding capacity, background control, sensitivity, and throughput. Use the quick criteria to narrow choices, then jump to featured SKUs for the most common use cases.

• Transfer Membranes (PVDF & Nitrocellulose) — PVDF offers higher protein binding and robustness for stripping/reprobing; nitrocellulose provides low background and easy handling. Choose 0.2 µm for small proteins and 0.45 µm for most targets. Representative Thermo Fisher lines include Pierce PVDF Transfer Membranes (e.g., 0.45 µm roll, Cat. 88518; 0.2 µm roll, Cat. 88520) and Nitrocellulose Membranes (e.g., 0.2 µm sheets, Cat. 88024; 0.45 µm roll, Cat. 88018).

• Protein Ladders/Markers — Use pre-stained standards to monitor transfer and orient blots quickly. PageRuler™ Prestained Protein Ladder, 10–180 kDa is a widely used choice (Cat. 26616, 26617).

• Transfer Stacks & Accessories — For fast, mess-free transfers, pre-assembled iBlot™ 2 Transfer Stacks in nitrocellulose (mini or regular) reduce setup time (Cat. IB23002, IB23001).

• Blocking & Wash Buffers — Match blocking chemistry to detection method (casein or BSA for HRP-ECL; low-fluorescence blockers for NIR). Maintain TBST strength and wash times to control background.

• Primary & Secondary Antibodies — Verify species, clonality, and conjugate (HRP/AP or fluorophores); filter for “WB-validated” when available.

• Chemiluminescent Substrates (ECL) — Pick sensitivity to fit protein abundance: SuperSignal™ West Pico PLUS for routine blots (Cat. 34580) and SuperSignal™ West Femto for low-abundance targets (Cat. 34094, 34096). 

• Stripping Buffers — For reprobing, Restore™ Western Blot Stripping Buffer offers multiple pack sizes (Cat. 21062, 21059, 21063). Use fluorescent-compatible strippers for NIR workflows.

Quick Selection Guide (Comparison Table)

Use this at-a-glance matrix to match your target and readout to a membrane, ECL level, and buffer set. It’s designed for fast triage; follow the links to product cards if you need specs or SDS/IFU files.

Featured Products (Validated Best-sellers)

Below are high-frequency picks our customers rely on. SKUs and formal names follow Thermo Fisher listings so your lab and purchasing team can order with confidence.

• SuperSignal™ West Pico PLUS Chemiluminescent Substrate, 500 mL — Cat. 34580. A balanced HRP substrate for routine western blots; supplied as Luminol/Enhancer (250 mL) + Stable Peroxide (250 mL). Good default for housekeeping and mid-abundance targets.

• SuperSignal™ West Femto Maximum Sensitivity Substrate — Cat. 34094 (options vary by pack size; also available as 34096). For low-abundance proteins, extended linear range helps minimize saturation in quantification.

• PageRuler™ Prestained Protein Ladder, 10–180 kDa — Cat. 26616 (2 × 250 µL), 26617 (10 × 250 µL). Ten colored bands for quick gel and transfer assessment; ready-to-load, no heating or reducing required.

• Pierce™ PVDF Transfer Membranes, 0.45 µm, roll — Cat. 88518. High binding capacity and durable handling; a versatile choice for general western blotting and multiple probing cycles.

• Pierce™ PVDF Transfer Membranes, 0.2 µm, roll — Cat. 88520. Optimized for small proteins/peptides to reduce “blow-through” during transfer.

• Nitrocellulose Membranes, 0.2 µm, 8 × 8 cm sheets — Cat. 88024. Low background, convenient sheets sized for mini-gels; a reliable routine option.

• Nitrocellulose Membranes, 0.45 µm, 3.5 m × 30 cm roll — Cat. 88018. Economical for high throughput; cut-to-fit for consistent lab stocking.

• iBlot™ 2 Transfer Stacks, nitrocellulose (mini/regular) — Cat. IB23002 (mini), IB23001 (regular). Pre-assembled stacks for rapid transfers with the iBlot 2 device; excellent for standardized, repeatable setups.

• Restore™ Western Blot Stripping Buffer — Cat. 21062 (30 mL), 21059 (500 mL), 21063 (5 L; also 21059X4). Efficient stripping for PVDF and nitrocellulose prior to reprobing.

Compatibility & Tips (Reduce Background, Improve Signal)

Most “mystery” issues are preventable with a few consistent checks. Use the pointers below to tighten your workflow without changing your core reagents.

• Membrane × Detection fit — Pair PVDF with femto-level ECL when you expect low abundance; use 0.2 µm for small proteins. Nitrocellulose often gives a lower background for routine blots and Pico-level ECL. 

• Blocking & washes — Start with casein/BSA at the vendor-recommended concentration, then tune TBST salt and Tween to background. Over-blocking or overly strong detergent can suppress the signal.

• Antibody dilutions — Re-optimize secondary dilutions when moving from Pico to Femto substrates to avoid overexposure artifacts; keep exposures in the linear range for densitometry.

• Transfer consistency — If corners fade or bands smear, verify stack assembly/pressure and equilibrate gels; pre-assembled stacks (iBlot 2) can reduce setup variability.

• Reprobe strategy — Strip gently first; if switching to fluorescence, choose fluorescent-compatible stripping buffers to avoid background.

Frequently Asked Questions

Below are concise answers to help you finalize selections and avoid common pitfalls. Each links back to relevant categories for details.

Q1: PVDF or nitrocellulose for my target?
For most medium-to-high MW targets, PVDF 0.45 µm or NC 0.45 µm both work; PVDF is sturdier for reprobing and high-sensitivity ECL, NC can yield lower background in routine blots. For small proteins, choose 0.2 µm membranes.

Q2: When do I need Femto-level ECL?
Use SuperSignal West Femto when your target is scarce or when you must shorten exposure times without sacrificing linearity. Stick with West Pico PLUS for typical validation and housekeeping. Re-titrate antibodies when changing sensitivity.

Q3: Can I strip and reprobe the same membrane?
Yes—Restore Stripping Buffer is designed for PVDF and NC. After stripping, re-block and verify the background before the next probe. For fluorescent detection later, use a fluorescent-compatible stripper.

Q4: Do pre-stained ladders affect Western detection?
No—PageRuler Prestained Ladder is compatible with both SDS-PAGE and western transfer/detection and provides visible bands during transfer and imaging.

Talk to Iright

Need help mapping your target and imaging system to a membrane/ECL combination? Contact Iright for fast quotations on Thermo Fisher western blot supplies, documentation packs (SDS/IFU/CoA), and bulk pricing. We can also pre-build a membrane + ECL + blocking bundle that matches your lab’s SOP.