Western blotting antibodies
Essential Loading Controls
Recommended housekeeping proteins for normalization. Choose controls with consistent expression in your sample type and consider subcellular localization (cytosolic, nuclear, membrane) and molecular weight separation.
| Target (Gene) | Expected size (kDa) | Compartment | Notes | Browse |
|---|---|---|---|---|
| β-Actin (ACTB) | ~42 | Cytoskeleton | Common for cytosolic proteins; can vary in differentiation or motility studies. | View ACTB antibodies |
| GAPDH (GAPDH) | ~36–38 | Cytosol | Beware metabolic modulation; good for many cell lines and tissues. | View GAPDH antibodies |
| α/β-Tubulin (TUBA/TUBB) | ~50–55 | Cytoskeleton | Useful when ACTB conflicts with target MW. | View Tubulin antibodies |
| Vinculin (VCL) | ~116 | Membrane/cytoskeleton | High MW control; good when separating mid/low MW targets. | View Vinculin antibodies |
| HPRT1 (HPRT1) | ~24 | Cytosol | Alternative housekeeping; useful in proliferation studies. | View HPRT1 antibodies |
| Lamin B1 (LMNB1) | ~66 | Nuclear | Normalization for nuclear fractions. | View Lamin B1 antibodies |
| Histone H3 (H3) | ~15–17 | Nuclear (chromatin) | For nuclear/chromatin preps; pair with H3K27me3 or H3K9ac as needed. | View Histone H3 antibodies |
| Na⁺/K⁺-ATPase (ATP1A1) | ~100–112 | Membrane | Membrane fraction control. | View ATP1A1 antibodies |
Tip: Also consider total protein stains (e.g., Ponceau S) as orthogonal normalization when housekeeping proteins vary.
Cancer Biology Targets
Frequently profiled markers in oncology WB panels. Where relevant, include both total and phospho-specific antibodies to read pathway activation and pair tumor suppressors with downstream targets.
| Target (Gene) | Expected size (kDa) | Notes | Browse |
|---|---|---|---|
| p53 (TP53) | ~53 | DNA damage response; tumor suppressor. Pair with MDM2, p21. | Browse p53 antibodies |
| Ki‑67 (MKI67) | ~320–360 | Proliferation marker; bands can appear diffuse at high MW. | Browse Ki‑67 antibodies |
| EGFR (ERBB1) | ~170 (p‑EGFR varies) | Receptor tyrosine kinase; check glycosylation states. | Browse EGFR antibodies |
| HER2 (ERBB2) | ~185 | Breast/gastric cancer marker; consider total & phospho. | Browse HER2 antibodies |
| AKT1/2/3 | ~56 (p‑AKT Ser473) | PI3K pathway. Include total and phospho‑AKT for signaling status. | Browse AKT antibodies |
| ERK1/2 (MAPK3/1) | 44/42 (p‑ERK1/2) | MAPK pathway readout; rapid phosphorylation dynamics. | Browse ERK antibodies |
Metabolism & Mitochondria
Combine glycolysis enzymes (HK2, PKM2, LDHA) with a complete OXPHOS panel (Complex I–V) to profile metabolic rewiring and mitochondrial function.
| Target (Gene) | Expected size (kDa) | Notes | Browse |
|---|---|---|---|
| Hexokinase 2 (HK2) | ~100 | Glycolysis; cancer metabolism marker. | HK2 antibodies |
| PKM2 (PKM) | ~58 | Warburg effect; dimer/monomer forms. | PKM2 antibodies |
| LDHA | ~36 | Lactate production; hypoxia‑responsive. | LDHA antibodies |
| NDUFB8 (Complex I) | ~20–25 | OXPHOS panel marker. | NDUFB8 antibodies |
| SDHB (Complex II) | ~30 | OXPHOS panel marker. | SDHB antibodies |
| UQCRC2 (Complex III) | ~48 | OXPHOS panel marker. | UQCRC2 antibodies |
| COX IV / COX4I1 (Complex IV) | ~17 | OXPHOS panel marker. | COX4I1 antibodies |
| ATP5A1 (Complex V) | ~55 | OXPHOS panel marker. | ATP5A1 antibodies |
Hypoxia & Oxidative Stress
Use HIF1A/CA9 for hypoxia readouts and NRF2 pathway markers (HMOX1, SOD2) for oxidative stress. Document O2 levels and treatment timing.
| Target (Gene) | Expected size (kDa) | Notes | Browse |
|---|---|---|---|
| HIF‑1α (HIF1A) | ~120 | Stabilized under low O2. | HIF1A antibodies |
| PHD2 (EGLN1) | ~46 | Hydroxylates HIF‑1α; hypoxia regulator. | EGLN1 antibodies |
| CA9 | ~54 | Hypoxia‑induced membrane enzyme. | CA9 antibodies |
| NRF2 (NFE2L2) | ~95–110 | Oxidative stress TF; band shifts with PTMs. | NRF2 antibodies |
| HO‑1 (HMOX1) | ~32 | Cytoprotective enzyme; induced by stress. | HO‑1 antibodies |
| MnSOD (SOD2) | ~25 | Mitochondrial antioxidant enzyme. | SOD2 antibodies |
DNA Damage & Repair
After genotoxic insult, track γH2AX with ATM/ATR signaling and homologous recombination proteins (RAD51, BRCA1) to assess repair pathway choice.
| Target (Gene) | Expected size (kDa) | Notes | Browse |
|---|---|---|---|
| γH2AX (H2AFX pS139) | ~15–17 | DSB marker; include proper nuclear extraction. | H2AX antibodies |
| ATM | ~350 | PIKK kinase; DNA damage signaling. | ATM antibodies |
| ATR | ~300 | Replication stress response. | ATR antibodies |
| BRCA1 | ~220 | HR repair; multiple bands possible. | BRCA1 antibodies |
| RAD51 | ~37 | Strand exchange factor; HR marker. | RAD51 antibodies |
| 53BP1 (TP53BP1) | ~213 | DSB repair choice marker. | 53BP1 antibodies |
ER Stress / Unfolded Protein Response
Profile the three UPR arms: PERK, IRE1α, and ATF6. BiP/GRP78 and CHOP provide robust ER stress readouts; watch for mobility shifts on phospho species.
| Target (Gene) | Expected size (kDa) | Notes | Browse |
|---|---|---|---|
| BiP / GRP78 (HSPA5) | ~78 | ER chaperone; canonical UPR marker. | HSPA5 antibodies |
| CHOP (DDIT3) | ~27 | UPR‑induced transcription factor. | CHOP antibodies |
| PERK (EIF2AK3) | ~125 | Phospho‑PERK mobility shift. | PERK antibodies |
| IRE1α (ERN1) | ~110 | RNase/kinase; XBP1 splicing. | IRE1 antibodies |
| ATF6 (full/cleaved) | ~90 / ~50 | Golgi cleavage produces active form. | ATF6 antibodies |
Ubiquitin–Proteasome & PTMs
Differentiate ubiquitin linkage types: K48 (degradation) vs K63 (signaling). Use denaturing lysis to resolve Ub ladders and include proteasome subunits as controls.
| Target (Gene) | Expected size (kDa) | Notes | Browse |
|---|---|---|---|
| Ubiquitin (K48 linkage) | Smear | Proteasome‑directed chains; laddering pattern. | K48 Ub antibodies |
| Ubiquitin (K63 linkage) | Smear | Signaling chains; non‑degradative. | K63 Ub antibodies |
| PSMB5 | ~26 | 20S core subunit; proteasome activity. | PSMB5 antibodies |
| Parkin (PRKN) | ~52 | E3 ligase; mitophagy. | Parkin antibodies |
| SUMO1 | ~11 | SUMOylation; appears as conjugate ladders. | SUMO1 antibodies |
Wnt/β‑Catenin & EMT
Assess Wnt activation via β‑catenin stabilization, then monitor EMT via E/N‑cadherin switch, Vimentin, and Snail for phenotypic transition.
| Target (Gene) | Expected size (kDa) | Notes | Browse |
|---|---|---|---|
| β‑Catenin (CTNNB1) | ~92 | Stabilization indicates Wnt activation. | CTNNB1 antibodies |
| GSK3β | ~46 | p‑Ser9 reflects inhibition. | GSK3β antibodies |
| AXIN1 | ~96 | Destruction complex component. | AXIN1 antibodies |
| E‑cadherin (CDH1) | ~120 | Loss indicates EMT. | CDH1 antibodies |
| N‑cadherin (CDH2) | ~130 | EMT gain marker. | CDH2 antibodies |
| Vimentin (VIM) | ~57 | Mesenchymal marker. | Vimentin antibodies |
| Snail (SNAI1) | ~29 | EMT transcription factor. | SNAI1 antibodies |
TGF‑β / SMAD Signaling
Phosphorylation of SMAD2/3 indicates active TGF‑β signaling. Track receptor levels (TGFBR1) and pro/active forms of TGFB1.
| Target (Gene) | Expected size (kDa) | Notes | Browse |
|---|---|---|---|
| SMAD2 (total / p‑) | ~52 | p‑Ser465/467 indicates activation. | SMAD2 antibodies |
| SMAD3 (total / p‑) | ~52–58 | p‑Ser423/425 indicates activation. | SMAD3 antibodies |
| SMAD4 | ~60 | Common mediator; tumor suppressor. | SMAD4 antibodies |
| TGFBR1 (ALK5) | ~55 | Receptor kinase; monitor levels. | TGFBR1 antibodies |
| TGF‑β1 (TGFB1) | ~44 (pro‑form) | Pro/active forms; check processing. | TGFB1 antibodies |
Notch / Hedgehog / Hippo
Use NICD fragments for Notch activation, GLI1/PTCH1 for Hedgehog, and YAP/TAZ (total vs p‑Ser127) for Hippo pathway status.
| Target (Gene) | Expected size (kDa) | Notes | Browse |
|---|---|---|---|
| Notch1 (full / NICD) | ~300 / ~110 | NICD indicates activation. | NOTCH1 antibodies |
| GLI1 | ~150–160 | Hedgehog transcription factor. | GLI1 antibodies |
| PTCH1 | ~160 | Hedgehog receptor; decreases when active. | PTCH1 antibodies |
| YAP1 (total / p‑Ser127) | ~65 | Hippo pathway; cytoplasmic when p‑. | YAP1 antibodies |
| TAZ (WWTR1) | ~43–50 | Hippo pathway co‑activator. | TAZ antibodies |
Angiogenesis & Endothelial
Pair ligand/receptor measurements: VEGFA with VEGFR2 (p‑Tyr1175). CD31 quantifies endothelial content; eNOS reflects vascular function.
| Target (Gene) | Expected size (kDa) | Notes | Browse |
|---|---|---|---|
| VEGF‑A (VEGFA) | ~19–25 | Secreted isoforms; reducing conditions recommended. | VEGFA antibodies |
| VEGFR2 (KDR) | ~230 | p‑Tyr1175 used for activation. | VEGFR2 antibodies |
| CD31 (PECAM1) | ~130 | Endothelial marker; glycosylation shifts MW. | CD31 antibodies |
| ANGPT2 | ~57 | Angiopoietin family; vascular remodeling. | ANGPT2 antibodies |
| eNOS (NOS3) | ~140 | Endothelial nitric oxide synthase. | eNOS antibodies |
Neurodegeneration
Complement APP processing with Tau (total/phospho) and α‑synuclein. Fractionation helps separate membrane and cytosolic pools.
| Target (Gene) | Expected size (kDa) | Notes | Browse |
|---|---|---|---|
| APP | ~100–130 | Multiple isoforms and processing fragments. | APP antibodies |
| Tau (MAPT; total / p‑) | ~50–60 | AT8 epitope common for p‑Tau WB. | Tau antibodies |
| α‑Synuclein (SNCA) | ~14–15 | Pre‑synaptic protein; monomer/oligomer forms. | SNCA antibodies |
| BACE1 | ~70 | β‑secretase in APP processing. | BACE1 antibodies |