Donkey anti-Rabbit IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed)

• Reactivity: Rb
• Applications: WB, Simple Western, ELISA, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Donkey
• Conjugate: HRP
• Format: Pre-adsorbed
• Concentration: 0.5 mg/ml

Donkey anti-Rabbit IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) Summary

• Immunogen: This Donkey anti-Rabbit IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) was developed against rabbit IgG-heavy and light chain.
• Specificity: By immunoelectrophoresis and ELISA this Donkey anti-Rabbit IgG (H+L) Secondary Antibody [HRP] (Pre-adsorbed) reacts specifically with rabbit IgG and with light chains common to other rabbit immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. Antiserum was cross adsorbed using bovine, chicken, goat, human, mouse, pig and rat immunosorbents to remove cross reactive Antibodies. Less than 0.1% cross reactivity to bovine, chicken, goat, horse, human, mouse, pig and rat IgG was detected. This antibody may cross react with IgG from other species.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Donkey
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:10000-1:100000
  • Immunocytochemistry/ Immunofluorescence 1:200 - 1:5000
  • Immunohistochemistry 1:200 - 1:5000
  • Immunohistochemistry-Paraffin 1:200 - 1:5000
  • Simple Western 1:50
  • Western Blot 1:5000 - 1:50000
• Application Notes: In Simple Western internal validation: Separated by size; antibody dilution of 1:50; matrix was 12-230 kDa; detected by Chemiluminescence.. See Simple Western Antibody Database for Simple Western validation: separated by size; antibody dilution of 1:50; matrix was 12-230 kDa; detected by Chemiluminescence.
• Readout System:
  • NovaLume Plus Chemiluminescent Reagent LUM-555

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: Phosphate Buffered Saline (PBS) containing 0.2% BSA
• Preservative: 0.05% Pro-Clean 400
• Concentration: 0.5 mg/ml
• Purity: Immunogen affinity purified

Background

Antibodies, also known as immunoglobulins (Igs) are critical for immunity and are grouped into five primary classes: IgG, IgM, IgA, IgD, and IgE. The most abundant antibody isotype is immunoglobulin G (IgG) with concentrations ranging from 7.5-22 mg/ml in human serum and has a molecular weight of 150 kDa. The major effector functions of IgG include neutralization, opsonization, complement fixation and antibody dependent cell-mediated cytotoxicity (ADCC). This monomeric immunoglobulin, expressed on the surface of mature B cells, is often depicted as a Y-shape and comprised of 2 heavy chains and 2 light chains linked by disulfide bonds. The heavy chain is type gamma including subtypes gamma 1, gamma 2, gamma 3, and gamma 4 while the light chain is either a kappa or lambda chain. An IgG molecule has two antigen binding sites, each consisting of a heavy and light chain N-terminal variable domain. When combined with the constant heavy chain 1 (Ch1) and the constant light chain domains, it forms the fragment antigen-binding (Fab) region (2 per antibody). The remaining domains (Ch2-Ch4) of both heavy chains make up the Fc region and contain a site for covalently linking an enzymatic or fluorochrome probe, such as HRP or Janelia Fluor 549, for target detection and visualization (1,2,3).

The 4 IgG subclasses, sharing 95% amino acid identity, include IgG1, IgG2, IgG3, and IgG4 for humans and IgG1, IgG2a, IgG2b, and IgG3 for mice. The relative abundance of each human subclass is 60% for IgG1, 32% for IgG2, 4% for IgG3, and 4% for IgG4. In an IgG deficiency, there may be a shortage of one or more subclasses (4).

References

1. Painter RH. (1998) Encyclopedia of Immunology (Second Edition). Elsevier. 1208-1211

2. Chapter 9 - Antibodies. (2012) Immunology for Pharmacy. Mosby 70-78

3. Schroeder H, Cavacini, L. (2010) Structure and Function of Immunoglobulins. J Allergy Clin Immunol. 125(2 0 2): S41-S52. PMID: 20176268

4. Vidarsson G, Dekkers G, Rispens T. (2014) IgG subclasses and allotypes: from structure to effector functions. Front Immunol. 5:520. PMID: 25368619

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Secondary Antibodies are guaranteed for 1 year from date of receipt.

Publications for IgG (H+L) Antibody (NB7185)(12)

Publications using NB7185

• Akuma M, Kim M, Zhu C et al. Loss of VHL-mediated pRb regulation promotes clear cell renal cell carcinoma Cell Death & Disease 2025-04-16 [PMID: 40240354]
• Reddy S, Sham R, Smith K et al. Immune checkpoint protein PD-L1 promotes transcription of angiogenic and oncogenic proteins IL-8, Bcl3 and STAT1 in ovarian cancer cells. The Journal of Biological Chemistry 2025-02-21 [PMID: 39988077]
• Wilson ER, Nunes GD, Shen S et Al. Loss of prohibitin 2 in Schwann cells dysregulates key transcription factors controlling developmental myelination Glia 2024-10-30 [PMID: 39215540]
• Reed CB, Frick LR, Weaver A et al. Deletion of Calcineurin in Schwann Cells Does Not Affect Developmental Myelination, But Reduces Autophagy and Delays Myelin Clearance after Peripheral Nerve Injury J. Neurosci. 2020-07-08 [PMID: 32641402] (Western Blot)
• Weinstock NI, Shin D, Dhimal N et al. Macrophages Expressing GALC Improve Peripheral Krabbe Disease by a Mechanism Independent of Cross-Correction Neuron 2020-04-28 [PMID: 32375064]
• Chae M, Son ED, Bae IH et al. UVB-dependent inhibition of lipin-1 protects against proinflammatory responses in human keratinocytes Exp Mol Med 2020-02-22 [PMID: 32080341] (IF/IHC)
• Dovmark TH, Saccomano M, Hulikova A et al. Connexin-43 channels are a pathway for discharging lactate from glycolytic pancreatic ductal adenocarcinoma cells Oncogene 2017-04-04 [PMID: 28368405] (WB)
• Fergani IFM Exploring the Role of SUMOylation During the Peripheral Nerve Development Thesis Jan 1 2021
• Cimenci C, Blackburn N, Sedlakova V Et al. Combined Methylglyoxal Scavenger and Collagen Hydrogel Therapy Prevents Adverse Remodeling and Improves Cardiac Function Post Myocardial Infarction Advanced Functional Materials 2021-10-12
• G Shacklefor, LN Marziali, Y Sasaki, A Claessens, C Ferri, NI Weinstock, AM Rossor, NJ Silvestri, ER Wilson, E Hurley, GJ Kidd, S Manohar, D Ding, RJ Salvi, ML Feltri, M D'Antonio, L Wrabetz A new mouse model of Charcot-Marie-Tooth 2J neuropathy replicates human axonopathy and suggest alteration in axo-glia communication PloS Genetics, 2022-11-09;18(11):e1010477. 2022-11-09 [PMID: 36350884] (WB)
• Jo S, Na H, Choi Y et al. Saponin attenuates diesel exhaust particle (DEP)-induced MUC5AC expression and pro-inflammatory cytokine upregulation via TLR4/TRIF/NF-kappa B signaling pathway in airway epithelium and ovalbumin (OVA)-sensitized mice Journal of Ginseng Research 2022-04-01 [PMID: 36312733] (WB)
• S Padmanabha, B Gaire, Y Zou, MM Uddin, I Vancurova IFNgamma-induced PD-L1 expression in ovarian cancer cells is regulated by JAK1, STAT1 and IRF1 signaling Cellular Signalling, 2022-07-09;97(0):110400. 2022-07-09 [PMID: 35820543] (WB)

Review for IgG (H+L) Antibody (NB7185) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Other.

reviewed by: Verified Customer WB Other 05/22/2015

Summary

• Application: Western Blot
• Sample Tested: cos7 whole cell lysate
• Species: Other
• Lot: A11