IGHG3 Antibody (OASA05856)

Catalog#: OASA05856

• Size: 1ml

• Description of Target: DONKEY ANTI SHEEP/GOAT IgG:HRP
• Gene Symbol: IGHG3
• Alias Symbols: IgG3; FLJ39988; FLJ40036; FLJ40253; FLJ40587; FLJ40789; FLJ40834; MGC45809; DKFZp686H11213; IGHG3
• Host: Donkey
• Protein Size (# AA): 476
• Specificity: IgG
• Application: IHC-AFF, ELISA, IHC-FFPE, WB
• Conjugation: HRP
• Immunogen: The immunogen for anti-IGHG3 antibody: sheep IgG
• Product Format: Purified IgG conjugated to Horseradish Peroxidase (HRP) - liquid
• Isotype: Polyclonal IgG
• Applications Info: Immunohistology - Frozen:    1/50 - 1/100
  ELISA:    1/12800 - 1/25600
  Western Blotting:    1/20000
  
  Preservative Stabilisers: 0.05% - Proclin^TM 300
  0.002% - Methylisothiazolone
  0.002% - Bromonitrodioxane
  
  Antiserum Preparation: Antisera to sheep IgG were raised by repeated immunisation of donkeys with highly purified antigen. Purified IgG was prepared by affinity chromatography.
  Approx Protein Conc: 1.0 mg/ml
  
  Buffer Solutions: Phosphate buffered saline pH7.2
• Protocol Information: Citation: 1: Hallström T, Resman F, Ristovski M, Riesbeck K. Binding of complement regulators to invasive nontypeable Haemophilus influenzae isolates is not increased compared to nasopharyngeal isolates, but serum resistance is linked to disease severity. J Clin Microbiol. 2010 Mar;48(3):921-7. Epub 2010 Jan 20. PubMed PMID: 20089757; PubMed Central PMCID: PMC2832458.
  Species: Normal Human Serum(NHS)
  Experiment Name: Serum binding assay
  Experiment Background: In the present study, the characteristics of invasive nontypeable Haemophilus influenzae (NTHi) infections, including evidence of immune deficiency in the individual patient and the clinical presentation of the septic event, were studied. Teresia et al. correlated these findings with the capacity to bind specific complement regulators and the in vitro serum resistance of the individual isolates.
  Experimental Steps: 1. To analyze whether NTHi from different isolation sites bound C4BP or factor H directly from NHS, bacteria were grown overnight in BHI broth. NTHi bacteria (109) were incubated with hiNHS and buffer (100 mM NaCl, 50 mM Tris-HCl, pH 7.4) for 1 h at37°C. 2. To remove unbound proteins, bacteria were washed 5 times with the same buffer. Thereafter, the bacterial pellet was resuspended in 150 µl of 0.1 M glycine-HCl, pH 2.0, in order to elute bound proteins.3. Bacteria without NHS were used as a negative control. After 15 min of incubation at 37°C with shaking, bacteria were centrifuged and the supernatants were subjected to SDS-PAGE (10%).4. To analyze whether NTHi bound vitronectin from NHS, bacteria were grown overnight and incubated with hiNHS and PBS. To removeunbound proteins, NTHi bacteria were washed 5 times with the same buffer.5. Thereafter, the bacterial pellet was resuspended in 50 µl of 0.1% Triton X-100 (Darmstadt, Germany) and protease inhibitors (Complete; Roche, Mannheim, Germany).6. After 30 min of incubation at 4°C, bacteria were centrifuged and the supernatants were subjected to SDS-PAGE (10%).7. Electrophoretic transfer of protein bands from the gel to an Immobilon-P membrane (Millipore, Bedford, MA) was done at 35 V for 2 h. After transfer, the Immobilon-P membrane was blocked in PBS with 0.1% Tween 20 (PBS-Tween) containing 5% milk powder. After several washings, the membrane was incubated with rabbit anti-human C4BP, goat anti-human factor H, or goat anti-human vitronectin pAb, followed by incubation with HRP-conjugated swine anti-rabbit or donkey-anti-goat pAb.8. After incubation and additional washings in PBS-Tween, development was performed with enhanced chemiluminescence (ECL) Western blotting detection reagents (Pierce, Rockford, IL).
  Other Reagents Used: NaCl, glucose, gelatin, MgCl2 , CaCl2.
  Number Of Protocols: 1
• Key Reference: 1. Singh, M. et al. (1999) A recombinant measles virus expressing hepatitis B virus surface antigen induces humoral immune responses in genetically modified mice. J. Virol. 73 (6): 4823 - 4828.
  2. Tedla, N. et al. (1998) Regulation of T lymphocyte trafficking into lymph nodes during an immune response by the chemokines macrophage inflammatory protein (MIP) - 1 alpha and MIP-1 beta. J. Immunol. 161: 5663 - 5672
  3. Turner, J. et al. (2002) in vivo IL-10 production reactivates chronic pulmonary tuberculosis in C57BL/6 mice. J. Immunol. 169: 6343 - 6351.
• Reconstitution and Storage: Store at +4^oC or at -20^oC if preferred.
  
  This product should be stored undiluted.
  
  Storage in frost free freezers is not recommended Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.

Product Protocols: Serum binding assay Protocol with IGHG3 Antibody (Catalog Number: OASA05856)

Product Protocols:
Serum binding assay Protocol with Gene Name: IGHG3 Antibody (Catalog Number: OASA05856)

Catalog Number: OASA05856

Reference:
1: Hallström T, Resman F, Ristovski M, Riesbeck K.Binding of complement regulators to invasive nontypeable Haemophilus influenzae isolates is not increased compared to nasopharyngeal isolates, but serum resistance is linked to disease severity.J Clin Microbiol.2010 Mar;48(3):921-7.Epub 2010 Jan 20.PubMed PMID: 20089757; PubMed Central PMCID: PMC2832458.

Product Name: IGHG3 Antibody (OASA05856)

Gene Name: IGHG3

Species: Sheep

Sample description: Defects in IGHG1 are a cause of multiple myeloma (MM) [MIM:254500].MM is a malignant tumor of plasma cells usually arising in the bone marrow and characterized by diffuse involvement of the skeletal system, hyperglobulinemia, Bence-Jones proteinuria and anemia.Complications of multiple myeloma are bone pain, hypercalcemia, renal failure and spinal cord compression.The aberrant antibodies that are produced lead to impaired humoral immunity and patients have a high prevalence of infection.Amyloidosis may develop in some patients.Multiple myeloma is part of a spectrum of diseases ranging from monoclonal gammopathy of unknown significance (MGUS) to plasma cell leukemia.Note=A chromosomal aberration involving IGHG1 is found in multiple myeloma.Translocation t(11;14)(q13;q32) with the IgH locus.Translocation t(11;14)(q13;q32) with CCND1; translocation t(4;14)(p16.3;q32.3) with FGFR3; translocation t(6;14)(p25;q32) with IRF4.

Experiment Name: Serum binding assay

Experiment Background:
To analyze whether NTHi from different isolation sites bound C4BP or factor H directly from NHS, bacteria were grown overnight in BHI broth.NTHi bacteria (109) were incubated with hiNHS and buffer (100 mM NaCl, 50 mM Tris-HCl, pH 7.4) for 1 h at37 C.2.To remove unbound proteins, bacteria were washed 5 times with the same buffer.Thereafter, the bacterial pellet was resuspended in 150 ul of 0.1 M glycine-HCl, pH 2.0, in order to elute bound proteins.
3.Bacteria without NHS were used as a negative control.After 15 min of incubation at 37 C with shaking, bacteria were centrifuged and the supernatants were subjected to SDS-PAGE (10%).
4.To analyze whether NTHi bound vitronectin from NHS, bacteria were grown overnight and incubated with hiNHS and PBS.To removeunbound proteins, NTHi bacteria were washed 5 times with the same buffer.
5.Thereafter, the bacterial pellet was resuspended in 50 ul of 0.1% Triton X-100 (Darmstadt, Germany) and protease inhibitors (Complete; Roche, Mannheim, Germany).
6.After 30 min of incubation at 4 C, bacteria were centrifuged and the supernatants were subjected to SDS-PAGE (10%).
7.Electrophoretic transfer of protein bands from the gel to an Immobilon-P membrane (Millipore, Bedford, MA) was done at 35 V for 2 h.After transfer, the Immobilon-P membrane was blocked in PBS with 0.1% Tween 20 (PBS-Tween) containing 5% milk powder.After several washings, the membrane was incubated with rabbit anti-human C4BP, goat anti-human factor H, or goat anti-human vitronectin pAb, followed by incubation with HRP-conjugated swine anti-rabbit or donkey-anti-goat pAb.
8.After incubation and additional washings in PBS-Tween, development was performed with enhanced chemiluminescence (ECL) Western blotting detection reagents (Pierce, Rockford, IL).
Experiment step:
In the present study, the characteristics of invasive nontypeable Haemophilus influenzae (NTHi) infections, including evidence of immune deficiency in the individual patient and the clinical presentation of the septic event, were studied.Teresia et al.correlated these findings with the capacity to bind specific complement regulators and the in vitro serum resistance of the individual isolates.