Goat anti-Rabbit IgG (Heavy & Light Chain) Antibody (FITC)

Details for Product No. ABIN101988, Supplier: Log in to see
Antigen
Epitope
Heavy & Light Chain
6012
1904
1868
1159
274
266
266
124
33
30
30
15
12
12
12
12
11
7
3
2
1
1
1
1
1
1
1
Reactivity
Rabbit
2740
2672
2191
1495
1370
700
664
447
434
362
359
354
275
258
257
147
115
84
43
26
24
21
12
12
8
7
5
5
4
4
4
3
3
3
2
2
2
2
2
2
1
1
1
Host
Goat
6467
4816
1755
887
458
274
87
54
42
35
22
8
6
5
4
1
1
1
Clonality
Polyclonal
Conjugate
FITC
2063
1740
1639
1144
678
644
422
315
171
122
102
102
94
89
88
88
86
84
79
78
74
73
70
68
68
67
65
65
63
55
55
54
54
54
54
54
53
53
53
53
53
48
47
47
41
40
32
31
30
23
23
22
20
19
18
17
16
15
15
15
14
14
14
12
12
12
12
10
10
10
10
10
9
9
9
9
8
8
8
8
8
7
6
6
6
6
6
6
6
5
5
5
5
5
5
5
5
5
5
4
4
4
4
3
3
3
3
3
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
1
1
1
1
1
1
1
Application
FLISA, Flow Cytometry (FACS), Fluorescence Microscopy (FM)
Options
Supplier
Log in to see
Supplier Product No.
Log in to see
'Independent Validation' Badge
Antigen Rabbit IgG (Heavy & Light Chain)
Lot Number 611-1202
Method validated Immunofluorescence
Positive Control Lab stock CBD-SNAP antibody
Negative Control No SNAP-tag antibody
Notes We validate the specificity of the secondary goat anti-rabbit IgG (heavy & light chain) antibody (FITC) ABIN101988 for rabbit IgG antibody.
Primary Antibody ABIN1573927
Secondary Antibody ABIN101988
Protocol
  • Oyster visceral mass tissue is dissected and fixed in 4% paraformaldehyde in seawater overnight.
  • Serial dehydration process using an automated ASP300S Enclosed Tissue Processor (Leica Biosystems) as follows:
    • 70% ethanol for 45min
    • 90% ethanol for 45min
    • 90% ethanol for 45min
    • 100% ethanol twice for 45min
    • xylene twice for 45min
    • paraffin wax at 58°C 3 times for 30 min
  • Tissue is mounted in a paraffin block and hardened overnight before.
  • 8µm tissue sections are retrieved from the block and collected on circular glass cover slips.
  • Heat cover slips at 60°C for 1h.
  • Deparaffination and rehydration:
    • Xylene twice for 15min
    • 100% ethanol twice for 10min
    • 95% ethanol for 10 min
    • 85% ethanol for 10 min
    • 70% ethanol for 10 min
    • 50% ethanol for 10 min
    • 30% ethanol for 10 min
    • distilled water for 10 min
    • PBS for 10 min
  • Wash tissue sections with PBS with 0.05% triton X twice for 30min.
  • Permeabilize in PBS with 0.05% triton X overnight.
  • Treatment of the tissue sections with 1mg/mL sodium borohydride in PBS three times for 5min to reduce autofluorescence.
  • Wash sections in PBS 3 times for 15 min for at RT.
  • Block sections in PBST with 1% BSA for 2 hours at RT.
  • Incubate sections with CBD-SNAP antibody (lab stock) diluted 1:200 in PBST with 1% BSA overnight at 4°C to detect the location of chitin.
  • Wash sections in PBS 3 times for 15min with PBS at RT.
  • Additionally, incubate the CBD-SNAP and SNAP-tag double-stained sections with rabbit anti-SNAP antibody (antibodies-online, ABIN1573927, lot 13D000621) diluted 1:200 in PBST with 1% BSA overnight at 4°C.
  • Wash sections in PBS 3 times for 15min with PBS at RT.
  • Incubate sections with the secondary goat anti-rabbit IgG (heavy & light chain) antibody (FITC) (antibodies-online, ABIN101988, lot 611-1202) diluted 1:400 in PBST with 1% BSA for 2h at °C.
  • Wash sections in PBS three times for 15min at RT.
  • Counterstain with 0.1µg/mL DAPI in PBS for 15min at RT.
  • Wash sections in PBS three times for 15min at RT.
  • Mount sections on a microscopic slide using 50% glycerol in PBS.
  • Seal cover slips with nail polish.
  • Confocal imaging on Leica SPE.
  • Visualization of the data performed on LAS 3D software.
Experimental Notes To validate the specificity of the anti-rabbit FITC secondary antibody ABIN101988, 8µm paraffin sections of oyster visceral mass were observed in this study. We compared the fluorescence signals with immunofluorescence study. The negative control specimen was always compared with the test specimen or the positive control specimen on the same day, using the same laser power, gain, offset, accumulation/averaging settings on the Leica SPE confocal microscope. Visualization of the data was performed on LAS 3D software, with the same visualization setting to compare signal brightness. We found that the samples treated with anti-rabbit FITC secondary antibody ABIN101988 had similar fluorescence signals as the positive control Anti Rabbit Alexa 488. Excitation at the same laser wavelength and power did not generate fluorescence in the negative control section, when anti-rabbit FITC secondary antibody was applied in the absence of rabbit produced anti-SNAP antibody.
Validation Images
Immunofluorescence validation image for Goat anti-Rabbit IgG (Heavy & Light Chain) antibody (FITC) (ABIN101988) Immunofluorescence images of oyster visceral mass tissue, with the specificity of Ant...
Immunogen

Immunogen: Rabbit IgG whole molecule

Isotype IgG
Specificity IgG (H&L)
Cross-Reactivity Rabbit
Characteristics Concentration Definition: by UV absorbance at 280 nm
Purification This product was prepared from monospecific antiserum by immunoaffinity chromatography using Rabbit IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities.  Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Fluorescein, anti-Goat Serum, Rabbit IgG and Rabbit Serum.
Labeling Ratio 3.1
Antigen
Background

Synonyms: Goat anti-Rabbit IgG Antibody fluorescein Conjugation, Goat anti-Rabbit IgG FITC Conjugated Antibody

Background: Anti-Rabbit IgG Antibody Fluorescein generated in goat detects rabbit IgG. Secreted as part of the adaptive immune response by plasma B cells, immunoglobulin G constitutes 75 % of serum immunoglobulins. Immunoglobulin G binds to viruses, bacteria, as well as fungi and facilitates their destruction or neutralization via agglutination (and thereby immobilizing them), activation of the compliment cascade, and opsinization for phagocytosis. The whole IgG molecule possesses both the F(c) region, recognized by high-affinity Fc receptor proteins, as well as the F(ab) region possessing the epitope-recognition site. Both heavy and light chains of the antibody molecule are present. Secondary Antibodies are available in a variety of formats and conjugate types. When choosing a secondary antibody product, consideration must be given to species and immunoglobulin specificity, conjugate type, fragment and chain specificity, level of cross-reactivity, and host-species source and fragment composition. This Anti-Rabbit IgG (H&L) is conjugated to Fluorescein.

Application Notes

Application Note: This product is designed for immunofluorescence microscopy, fluorescence based plate assays (FLISA) and fluorescent western blotting. This product is also suitable for multiplex analysis, including multicolor imaging, utilizing various commercial platforms.

FLISA Dilution: 1:10,000 - 1:50,000

Flow Cytometry Dilution: 1:500 - 1:2,500

IF Microscopy Dilution: 1:1,000 - 1:5,000

Comment

Excitation/Emission wavelength: 494 nm/514 nm

Restrictions For Research Use only
Format Lyophilized
Reconstitution

Reconstitution Volume: 1.0 mL

Reconstitution Buffer: Restore with deionized water (or equivalent)

Concentration 2.0 mg/mL
Buffer

Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Stabilizer: 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free

Preservative: 0.01 % (w/v) Sodium Azide

Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Product is photosensitive and should be protected from light.
Storage RT,4 °C,-20 °C
Expiry Date 12 months
Supplier Images
Dot Blot (DB) image for Goat anti-Rabbit IgG (Heavy & Light Chain) antibody (FITC) (ABIN101988) FITC (fluorescein) and HRP (horse radish peroxidase) conjugated secondary antibody wa...
Western Blotting (WB) image for Goat anti-Rabbit IgG (Heavy & Light Chain) antibody (FITC) (ABIN101988) Western Blot of Anti-Rabbit IgG (H&L) (GOAT) Antibody . Lane M: 3 μl Molecular Ladder...
Western Blotting (WB) image for Goat anti-Rabbit IgG (Heavy & Light Chain) antibody (FITC) (ABIN101988) Western blot of Fluorescein conjugated Goat Anti-Rabbit IgG secondary antibody. Lane ...
Product cited in: Krawiec, Liao, Kwan, DAmore, Weinbaum, Rubin, Wagner, Vorp: "Evaluation of the stromal vascular fraction of adipose tissue as the basis for a stem cell-based tissue-engineered vascular graft." in: Journal of vascular surgery, Vol. 66, Issue 3, pp. 883-890.e1, 2017 (PubMed).

Krawiec, Weinbaum, Liao, Ramaswamy, Pezzone, Josowitz, DAmore, Rubin, Wagner, Vorp: "In Vivo Functional Evaluation of Tissue-Engineered Vascular Grafts Fabricated Using Human Adipose-Derived Stem Cells from High Cardiovascular Risk Populations." in: Tissue engineering. Part A, Vol. 22, Issue 9-10, pp. 765-75, 2017 (PubMed).

Jiang, Xia, Yang, Shao, Liao, Zhu, Jiang: "Novel insights into a treatment for aplastic anemia based on the advanced proliferation of bone marrow‑derived mesenchymal stem cells induced by fibroblast growth factor 1." in: Molecular medicine reports, Vol. 12, Issue 6, pp. 7877-82, 2016 (PubMed).

Xu, Xu, Wang, Zhang: "Lefty1 alleviates renal tubulointerstitial injury in mice with unilateral ureteral obstruction." in: Molecular medicine reports, Vol. 13, Issue 1, pp. 901-8, 2016 (PubMed).

Background publications Blakeslee, Baines: "Immunofluorescence using dichlorotriazinylaminofluorescein (DTAF). I. Preparation and fractionation of labelled IgG." in: Journal of immunological methods, Vol. 13, Issue 3-4, pp. 305-20, 1977 (PubMed).