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- Table of Contents
IGKC / Immunoglobulin kappa constant · Western blot design guide
Design a Western Blot for IGKC
Real validated IGKC Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-IGKC WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
IGKC Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~11.8 kDa | |
| Gel | 15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Disulfide-linked | |
| Caveat | Reducing conditions | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated IGKC Western Blot Protocols
The M05470-5 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Human fetal spleen lysate (catalog M05470-5) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M05470-5; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected IGKC Western Blot Band Size?
IGKC is predicted at 11.8 kDa; disulfide-linked immunoglobulin may migrate differently, but its band position has not been demonstrated here.
| Band near 11.8 kDa | Consistent with the predicted isolated IGKC sequence; confirm identity with band controls |
| Higher band under nonreducing conditions | May reflect disulfide-linked light and heavy chains |
| Band shifts after reduction | Disrupting the documented disulfide bonds may change migration |
| Little or no band in whole-cell lysate | IGKC is also secreted, so some protein may be outside the cells |
💡Expected IGKC appearanceUniProt predicts 11.8 kDa for the IGKC sequence; disulfide-linked immunoglobulin may migrate differently, so use reducing and nonreducing lanes and an appropriate positive control to assess band identity.
| UniProt predicted mass | The 107-residue IGKC sequence has a predicted mass of 11.8 kDa |
| Cys27–Cys87 intrachain disulfide | Reduction may alter migration without changing sequence mass |
| Cys107 interchain disulfide | Can retain IGKC in a larger linked species under nonreducing conditions |
| Disulfide-linked heavy and light chains | An intact immunoglobulin complex can migrate above the isolated IGKC sequence |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | IGKC can be secreted | Check the culture supernatant alongside lysate and include a positive control |
| Band higher than expected | Disulfide-linked heavy and light chains may remain together | Compare reducing and nonreducing lanes |
| Band lower than expected | Proteolysis may have produced a smaller antibody-reactive fragment | Use fresh samples with protease inhibitors and confirm identity with an independent antibody |
| Multiple bands | Linked and reduced immunoglobulin species may coexist | Compare fully reduced and nonreducing samples |
| Weak or no signal | The tested sample may contain little IGKC | Include an IGKC-positive sample and examine the secreted fraction |
| Fragments below expected size | Sample proteolysis may produce antibody-reactive fragments | Minimize handling time, add protease inhibitors, and compare fresh samples |
Experiment evidence
Western blot images showing IGKC band
M05470-5 · IGKC
M05470-1 · IGKC
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for IGKC Western blot
🧪For positive controls for IGKC in Western blot, you can use no HPA-supported positive sample from the supplied evidence.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: IGKC is secreted, so whole-cell lysate may give a weak signal; a positive control cannot be selected from the supplied HPA data.
HPA tissue expression evidence for IGKC
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced IGKC Western Blot Tips
Deeper troubleshooting and optimisation questions for IGKC, answered from its protein features.
How should IGKC band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Do annotated IGKC isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands; assess sample composition and reducing conditions before assigning them.
Is a glycosylation-related IGKC shift supported?
PTM · No glycosylation site or modified residue is listed for this IGKC sequence. The supplied features do not support attributing a band shift to an annotated modification, and they do not establish whether any shift will be visible.
Does this guide establish induction of IGKC?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
How should transfer be checked for IGKC?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M05470-5 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should IGKC be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Why might an IGKC band differ from 11.8 kDa?
Interpretation · The 11.8 kDa prediction applies to the 107-residue constant region. Immunoglobulin light chains also contain a variable region and are disulfide-linked to heavy chains. Interpret band size in light of the sample and whether the blot was run under reducing conditions; these features alone do not establish an apparent band size.
Immunoglobulins contain two identical heavy chains and two identical light chains linked by disulfide bonds; IGKC has two annotated disulfide bonds. Compare reducing and nonreducing lanes when assessing whether a higher-mass signal could reflect linked chains.
IGKC is annotated as secreted and associated with the cell membrane. Consider both extracellular and membrane-containing samples when choosing a fraction; compare like fractions when interpreting band intensity.
Keep the sampled fraction and reducing conditions consistent across lanes. IGKC can occur in secreted and membrane-associated immunoglobulins, and disulfide-linked chains may affect which species a band represents.
Consider disulfide-linked immunoglobulin chains, especially in a nonreducing lane. The supplied features support that possibility but do not identify any observed band; a reducing comparison can help evaluate it.
Immunoglobulins contain two identical heavy chains and two identical light chains linked by disulfide bonds; IGKC has two annotated disulfide bonds. Compare reducing and nonreducing lanes when assessing whether a higher-mass signal could reflect linked chains.
IGKC is annotated as secreted and associated with the cell membrane. Consider both extracellular and membrane-containing samples when choosing a fraction; compare like fractions when interpreting band intensity.
Keep the sampled fraction and reducing conditions consistent across lanes. IGKC can occur in secreted and membrane-associated immunoglobulins, and disulfide-linked chains may affect which species a band represents.
Consider disulfide-linked immunoglobulin chains, especially in a nonreducing lane. The supplied features support that possibility but do not identify any observed band; a reducing comparison can help evaluate it.
Boster reagents
IGKC Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-Human Kappa Chain Rabbit Monoclonal Antibody
Cat # M05470-5
Real WB data
Anti-Kappa light chain IGKC Rabbit Monoclonal Antibody
Cat # M05470-1
Two the supplier rabbit monoclonal anti-IGKC antibodies are listed with human reactivity and Western blot images. The supplied evidence shows M05470-5 tested on human fetal spleen lysate and M05470-1 on human plasma lysate; performance in other samples is not established here.
Which to pick: For human fetal spleen lysate, consider M05470-5; for human plasma lysate, consider M05470-1. Both have Western blot images. Choose based on the tested sample context, since the supplied evidence does not establish broader performance.
Source: BosterBio IGKC gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P01834.
- Human Protein Atlas. IGKC tissue expression.
- PMC2965569 — target-verified WB comparison
- PMC5816075 — target-verified WB comparison
- PMC4848925 — target-verified WB comparison