IHC (Immunohistochemistry)
(CD45 (leukocyte common antigen, LCA) expression in lymphoid cells of the palatine tonsil (A) and lymph node(B). Formalin fixed, paraffin embedded human tissues(4 um sections) stained with anti - CD45 (AAA14929) monospecific clonal antibody according to related datasheet.)
Rabbit CD 45 Monoclonal Antibody | anti-CD 45 antibody
Anti - CD 45
Gene Names
PTPRC; LCA; LY5; B220; CD45; L-CA; T200; CD45R; GP180
Applications
Immunohistochemistry
Synonyms
CD 45, Antibody; Anti - CD 45; anti-CD 45 antibody
Host
Rabbit
Clonality
Monoclonal
Clone Number
E19-G
Specificity
Human
Form/Format
20 mM Tris-HCI, pH 8.0
Sequence Length
1304
Applicable Applications for anti-CD 45 antibody
Immunohistochemistry (IHC-P)-Paraffin
Application Notes
IHC-P dilution 1:100
IHC-P PROTOCOL-INSTRUCTION MANUAL:
1. Deparaffinize the section in 3 changes of xylene, 5 minutes each.
2. Wash the section in 96%, 80% and 70% ethyl alcohol for 10 minutes each.
3. Rinse in distilled water.
4. Block the endogenous peroxidase by incubating the tissue in 3% hydrogen peroxide (H2O2) for 10 minutes.
5. Wash in distilled water for 5 minutes.
6. For antigen retrieval: immerse the slide in the citrate buffer, pH 6.0, 0.05% Tween-20*, and incubate in water bath at 96-98 degree C for 30-40 minutes. (Alternatively adjust to your own protocol, keeping the required pH).
7. Transfer the slide to room temperature and let it cool down (in citrate buffer, pH 6.0) for 15 minutes.
8. Rinse in distilled water.
9. Wash in 0.05 M Tris-HCl, pH 7.6 buffer supplemented with 0.2% of Tween-20 (buffer A) for 5 minutes.
10. CONCENTRATED:
Incubate the section with primary antibody at the dilution 1:100 for 1 hour in the closed wet chamber.
READY TO USE (RTU):
Incubate the section with primary antibody (ready to use) for 1 hour in a closed wet chamber.
11. Wash twice 5 minutes with buffer A.
12. Apply the secondary antibody (the protocol depends on the supplier), and proceed to standard immunohistochemistry protocol (HRP - Peroxide - DAB). Micropolymer-HRP detection kit rabbit/mouse dual is suggested.
13. Wash twice 5 minutes with buffer A.
14. Apply the chromogen (DAB), 1 - 3 minutes.
15. Wash in water for 10 minutes.
16. Stain in hematoxylin for 5 minutes.
17. Wash in water for 10 minutes.
18. Dehydrate the section in 2 changes of 96% ethyl alcohol for 5 minutes each.
19. Wash the section in 2 changes of xylene for 2 minutes each.
20. Mount the slide for observation.
* Citrate Buffer (10mM Citric Acid, 0.05% Tween-20, pH 6.0):
Citric acid (anhydrous) ---------- 1.92 g; Distilled water ----------- 1000 ml
Mix to dissolve in 700 ml of distilled water.
Adjust pH to 6.0 with 1M NaOH and then add 0.5 ml of Tween-20 and mix well.
Adjust the final volume to 1 liter with distilled water.
Store this solution at room temperature for 3 months or at +4°C for longer storage.
IHC-P PROTOCOL-INSTRUCTION MANUAL:
1. Deparaffinize the section in 3 changes of xylene, 5 minutes each.
2. Wash the section in 96%, 80% and 70% ethyl alcohol for 10 minutes each.
3. Rinse in distilled water.
4. Block the endogenous peroxidase by incubating the tissue in 3% hydrogen peroxide (H2O2) for 10 minutes.
5. Wash in distilled water for 5 minutes.
6. For antigen retrieval: immerse the slide in the citrate buffer, pH 6.0, 0.05% Tween-20*, and incubate in water bath at 96-98 degree C for 30-40 minutes. (Alternatively adjust to your own protocol, keeping the required pH).
7. Transfer the slide to room temperature and let it cool down (in citrate buffer, pH 6.0) for 15 minutes.
8. Rinse in distilled water.
9. Wash in 0.05 M Tris-HCl, pH 7.6 buffer supplemented with 0.2% of Tween-20 (buffer A) for 5 minutes.
10. CONCENTRATED:
Incubate the section with primary antibody at the dilution 1:100 for 1 hour in the closed wet chamber.
READY TO USE (RTU):
Incubate the section with primary antibody (ready to use) for 1 hour in a closed wet chamber.
11. Wash twice 5 minutes with buffer A.
12. Apply the secondary antibody (the protocol depends on the supplier), and proceed to standard immunohistochemistry protocol (HRP - Peroxide - DAB). Micropolymer-HRP detection kit rabbit/mouse dual is suggested.
13. Wash twice 5 minutes with buffer A.
14. Apply the chromogen (DAB), 1 - 3 minutes.
15. Wash in water for 10 minutes.
16. Stain in hematoxylin for 5 minutes.
17. Wash in water for 10 minutes.
18. Dehydrate the section in 2 changes of 96% ethyl alcohol for 5 minutes each.
19. Wash the section in 2 changes of xylene for 2 minutes each.
20. Mount the slide for observation.
* Citrate Buffer (10mM Citric Acid, 0.05% Tween-20, pH 6.0):
Citric acid (anhydrous) ---------- 1.92 g; Distilled water ----------- 1000 ml
Mix to dissolve in 700 ml of distilled water.
Adjust pH to 6.0 with 1M NaOH and then add 0.5 ml of Tween-20 and mix well.
Adjust the final volume to 1 liter with distilled water.
Store this solution at room temperature for 3 months or at +4°C for longer storage.
Stabilizer
20 mg/ml BSA
Preservative
0.05% NaN3
Immunogen
Peptide derived from C-terminal sequence of human CD45. Antibody recognizes the epitope between Glu1277 - Val1292.
Cellular localization
Membrane
Positive control
Human tonsil tissue
VENTANA PROTOCOL-INSTRUCTION MANUAL
SHORT APPLICATION PROTOCOL FOR VENTANA BENCHMARK SLIDE STAINING SYSTEM
PROCEDURE: U ultraView DAB
1. Deparafinization
2. Heating (72 degree C) at the medium temperatures. Deparafinization.
3. Cell conditioning
4. ULTRA conditioner #2
5. Heating glass (95 degree C), incubation 8 min. (Cell conditioner #2; buffer CC2).
6. ULTRA CC2 solution application – 44 min.
7. Antibody incubation temperature
8. Heating glass (36 degree C), incubation 4 min.
9. Titration
10. Hand apply – primary antibody 100 ul. Incubation 32 min.
11. ultraWash.
12. Nuclear stain.
13. Hematoxylin II application – one drop (nuclear stain). Cover and incubate 12 min.
14. After nuclear stain.
15. Bluing reagent application, one drop. After nuclear stain, cover and incubate 4 min.
PROCEDURE: U ultraView DAB
1. Deparafinization
2. Heating (72 degree C) at the medium temperatures. Deparafinization.
3. Cell conditioning
4. ULTRA conditioner #2
5. Heating glass (95 degree C), incubation 8 min. (Cell conditioner #2; buffer CC2).
6. ULTRA CC2 solution application – 44 min.
7. Antibody incubation temperature
8. Heating glass (36 degree C), incubation 4 min.
9. Titration
10. Hand apply – primary antibody 100 ul. Incubation 32 min.
11. ultraWash.
12. Nuclear stain.
13. Hematoxylin II application – one drop (nuclear stain). Cover and incubate 12 min.
14. After nuclear stain.
15. Bluing reagent application, one drop. After nuclear stain, cover and incubate 4 min.
LEICA BOND MAX PROTOCOL-INSTRUCTION MANUAL
SHORT APPLICATION PROTOCOL FOR LEICA BOND MAX SLIDE STAINING SYSTEM
Protocol F:
- Visualization system: BOND Refine DS9800
- Epitope retrieval / heating time / temperature: ER1 / 30 min. / 100 degree C
- Incubation of primary antibody / temperature: 30 min. / 20 degree C
Protocol F:
- Visualization system: BOND Refine DS9800
- Epitope retrieval / heating time / temperature: ER1 / 30 min. / 100 degree C
- Incubation of primary antibody / temperature: 30 min. / 20 degree C
PRECAUTIONS
1. We strongly recommend to use Primary Antibody Diluent (catalog number please inquire), eventually alternative diluent (containing protease free BSA at the concentrations > 1mg/ml) for dilution of concentrated antibodies, otherwise the warranty might be voided.
2. Centrifuge the vial before use.
3. Intended for research use in laboratories.
4. Do not use after expiration date stamped on vial label.
5. Avoid contamination of the reagent.
6. Any discrepancies in the recommended procedures stated in the working protocol may affect the final results.
7. The reagent contains sodium azide (NaN3) which is highly toxic in higher concentrations. The concentration in the reagent (0.05%) is not considered as hazardous.
8. Disposal of waste material must be conducted in accordance with local regulations.
9. Wear appropriate Personal Protective Equipment to avoid contact with eyes and skin.
2. Centrifuge the vial before use.
3. Intended for research use in laboratories.
4. Do not use after expiration date stamped on vial label.
5. Avoid contamination of the reagent.
6. Any discrepancies in the recommended procedures stated in the working protocol may affect the final results.
7. The reagent contains sodium azide (NaN3) which is highly toxic in higher concentrations. The concentration in the reagent (0.05%) is not considered as hazardous.
8. Disposal of waste material must be conducted in accordance with local regulations.
9. Wear appropriate Personal Protective Equipment to avoid contact with eyes and skin.
Preparation and Storage
Store at +4 degrees C. Do not freeze.
IHC (Immunohistochemistry)
(CD45 (leukocyte common antigen, LCA) expression in lymphoid cells of the palatine tonsil (A) and lymph node(B). Formalin fixed, paraffin embedded human tissues(4 um sections) stained with anti - CD45 (AAA14929) monospecific clonal antibody according to related datasheet.)
IHC (Immunohistochemistry)
(CD45 (leukocyte common antigen, LCA) expression in lymphoid cells of the palatine tonsil (A) and lymph node(B). Formalin fixed, paraffin embedded human tissues(4 um sections) stained with anti - CD45 (AAA14929) monospecific clonal antibody according to related datasheet.)
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
NCBI Official Full Name
Receptor-type tyrosine-protein phosphatase C
NCBI Official Synonym Full Names
protein tyrosine phosphatase, receptor type, C
NCBI Official Symbol
PTPRC
NCBI Official Synonym Symbols
LCA; LY5; B220; CD45; L-CA; T200; CD45R; GP180
NCBI Protein Information
receptor-type tyrosine-protein phosphatase C; CD45 antigen; T200 glycoprotein; T200 leukocyte common antigen; protein tyrosine phosphatase, receptor type, c polypeptide
UniProt Protein Name
Receptor-type tyrosine-protein phosphatase C
UniProt Gene Name
PTPRC
UniProt Synonym Gene Names
CD45; L-CA
UniProt Entry Name
PTPRC_HUMAN
Similar Products
Product Notes
The CD 45 ptprc (Catalog #AAA14929) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's CD 45 can be used in a range of immunoassay formats including, but not limited to, Immunohistochemistry (IHC-P)-Paraffin. IHC-P dilution 1:100 IHC-P PROTOCOL-INSTRUCTION MANUAL: 1. Deparaffinize the section in 3 changes of xylene, 5 minutes each. 2. Wash the section in 96%, 80% and 70% ethyl alcohol for 10 minutes each. 3. Rinse in distilled water. 4. Block the endogenous peroxidase by incubating the tissue in 3% hydrogen peroxide (H2O2) for 10 minutes. 5. Wash in distilled water for 5 minutes. 6. For antigen retrieval: immerse the slide in the citrate buffer, pH 6.0, 0.05% Tween-20*, and incubate in water bath at 96-98 degree C for 30-40 minutes. (Alternatively adjust to your own protocol, keeping the required pH). 7. Transfer the slide to room temperature and let it cool down (in citrate buffer, pH 6.0) for 15 minutes. 8. Rinse in distilled water. 9. Wash in 0.05 M Tris-HCl, pH 7.6 buffer supplemented with 0.2% of Tween-20 (buffer A) for 5 minutes. 10. CONCENTRATED: Incubate the section with primary antibody at the dilution 1:100 for 1 hour in the closed wet chamber. READY TO USE (RTU): Incubate the section with primary antibody (ready to use) for 1 hour in a closed wet chamber. 11. Wash twice 5 minutes with buffer A. 12. Apply the secondary antibody (the protocol depends on the supplier), and proceed to standard immunohistochemistry protocol (HRP - Peroxide - DAB). Micropolymer-HRP detection kit rabbit/mouse dual is suggested. 13. Wash twice 5 minutes with buffer A. 14. Apply the chromogen (DAB), 1 - 3 minutes. 15. Wash in water for 10 minutes. 16. Stain in hematoxylin for 5 minutes. 17. Wash in water for 10 minutes. 18. Dehydrate the section in 2 changes of 96% ethyl alcohol for 5 minutes each. 19. Wash the section in 2 changes of xylene for 2 minutes each. 20. Mount the slide for observation. * Citrate Buffer (10mM Citric Acid, 0.05% Tween-20, pH 6.0): Citric acid (anhydrous) ---------- 1.92 g; Distilled water ----------- 1000 ml Mix to dissolve in 700 ml of distilled water. Adjust pH to 6.0 with 1M NaOH and then add 0.5 ml of Tween-20 and mix well. Adjust the final volume to 1 liter with distilled water. Store this solution at room temperature for 3 months or at +4°C for longer storage. Researchers should empirically determine the suitability of the CD 45 ptprc for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "CD 45, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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