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R7757

Sigma-Aldrich

Red Blood Cell Lysing Buffer Hybri-Max™

Liquid, sterile-filtered, suitable for hybridoma

Synonym(s):

RBC Lysing Buffer

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About This Item

UNSPSC Code:
12352207
NACRES:
NA.75

Product Name

Red Blood Cell Lysing Buffer Hybri-Max™, liquid, sterile-filtered, suitable for hybridoma

sterility

sterile-filtered

Quality Level

400

form

liquid

technique(s)

cell culture | hybridoma: suitable

impurities

endotoxin, tested

Application

Recommended for use by adding 1 mL of buffer to a cell pellet (cell pellet = 1 spleen or 100-200 million cells). Gently mix for 1 minute. Dilute the buffer with 15-20 mL of medium or salt solution. Centrifuge at 250-500 × g for 7 minutes and decant the supernatant. Cells may be diluted and prepared for counting or fusion. If lysis is incomplete, steps 1-4 may be repeated.

Biochem/physiol Actions

Red Blood Cell Lysing Buffer has been developed for use in hybridoma protocols to remove red blood cells from mouse splenocyte suspensions before fusion. It is also useful in systems where it may be desirable to remove red blood cells from cell suspensions, such as whole blood.

Components

Contains 8.3 g/L ammonium chloride in 0.01 M Tris-HCl buffer.

Other Notes

Note: This product is intended for the removal of red blood cells from mice. This product may not be appropriate for the lysis of red blood cells of other animals. The suitability of the product in any application other than mouse splenocytes must be determined by the researcher.

Legal Information

Hybri-Max is a trademark of Sigma-Aldrich Co. LLC

Storage Class Code

12 - Non Combustible Liquids

WGK

nwg

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

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Certificates of Analysis (COA)

Lot/Batch Number
RNBM0102
RNBL8380
RNBL6009
RNBL3445
RNBL1226

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  1. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  2. Is the BSA standard supplied with Product B6916, Bradford Reagent?

    No, we do not supply the BSA standard with this reagent. We recommend using one of the following products: Protein Standard (BSA) Solution, (2 mg/mL), Product No. P0834 or Protein Standard (BSA) Solution, (1 mg/mL), Product No. P0914, if lower concentrations of protein are to be measured.

  3. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  4. How do I find price and availability?

    There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers.

  5. What is the Department of Transportation shipping information for this product?

    Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product. 

  6. Can Product R7757, Red Blood Cell Lysing Buffer Hybri-Max™, be used to lyse cells that are trapped in whole tissue preparations?

    Unfortunately, we have not verified the suitability of this lysis buffer for using with tissue samples and do not have a protocol.   

  7. In step 3 of your usage sheet for Product R7757, Red Blood Cell Lysing Buffer Hybri-Max™, you say to dilute this buffer with medium or "salt solution". What salt solution is appropriate?

    Any balanced salt solution such as DPBS (Dulbecco's phosphate buffered saline) or HBSS (Hank's Balanced Salt Solution) may be used to dilute the buffer when following this procedure.

  8. Since you do not recommend Product R7757, Red Blood Cell Lysing Buffer Hybri-Max™, for treating whole blood, what protocol and products do you recommend for red cell lysis in a sample of whole blood?

    The following is a suggested protocol:1. Centrifuge to remove plasma. A volume of 0.2 mL of whole blood will yield approximately 0.1 mL of packed cells. Add 0.1ml of 1X buffer such as HBSS (Hanks' balanced salt solution, Product No. H1641) or DPBS (Dulbecco's phosphate buffered saline solution, Product No. D1283), to 0.1 mL of the pelleted cells and resuspend the pellet.2. Add 1-2 mls of 0.1X diluent (or water) and quickly mix cells using the same pipette.3. After 15 seconds, quickly add a volume of 2X diluent in the amount used in step 2 and mix thoroughly.  Dilute further with 1X diluent.  Wash cells once with buffer.

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